CN1890235A

CN1890235A - Benzopyranone compounds, compositions thereof, and methods of treatment therewith

Info

Publication number: CN1890235A
Application number: CNA2004800331210A
Authority: CN
Inventors: 杰弗里·A·麦金; 施拉佩得·S·巴格瓦; 约翰尼·贝纳乌德; 马丁·米斯拜克
Original assignee: Novartis AG; Signal Pharmaceuticals LLC
Current assignee: Novartis AG; Signal Pharmaceuticals LLC
Priority date: 2003-09-15
Filing date: 2004-09-13
Publication date: 2007-01-03
Also published as: US20050137231A1; US7247646B2; CA2538874A1; EP1667999A4; WO2005028472A1; AU2004274445A1; EP1667999A1; JP2007505835A; BRPI0414386A; MXPA06002876A

Abstract

Benzopyranone compounds having the following structure: wherein R1, R2, X, Y and n are as defined herein, are disclosed. The benzopyranone compounds are useful for treating or preventing a bone-resorbing disease, a neoplastic disease, arthritis, a disease exacerbated by the presence of estrogen or a disease improved by the presence of estrogen, comprising administering an effective amount of a benzopyranone compound to a patient in need of the treating or preventing; activating the function of estrogen receptor (''ER'') in a bone cell; inhibiting the function of ER in a cancer cell; inhibiting the expression of interleukin-6 (''IL-6'') in a cell; and inhibiting the growth of a neoplastic cell, comprising contacting the cell with an effective amount of a benzopyranone compound.

Description

Chromone compound, its composition and with its method for the treatment of

The application requires the right of priority of the U.S. Provisional Application submitted on September 15th, 2003 number 60/503,295, and this application is included in this paper as a reference in full.

1. invention field

The present invention relates to chromone compound, the composition that contains the chromone compound of significant quantity, and the method for treatment or prevention bone-resorbing disease, neoplastic disease, sacroiliitis, the disease that when oestrogenic hormon exists, worsens or the disease when oestrogenic hormon exists, improved, this method comprises the chromone compound that the patient of these needs significant quantity is arranged; Also relate to the function that activates estrogen receptor (" ER ") in the osteocyte, the function of ER in the anticancer, suppress the expression of interleukin-6 (" IL-6 ") in the cell, and the method that suppresses growth of tumour cell, this method comprises the chromone compound that makes the cells contacting significant quantity.

2. background of invention

Oestrogenic hormon has many effects for the women and the male sex's tissue.These biological action majorities are positive, comprise that keeping bone density, cardiovascular protection, central nervous system (" CNS ") function and armour system not influenced by aging.Yet except its positive effect, oestrogenic hormon still is the mammary gland and the endometrial potential growth factor, and this increases cancered risk.

Up to date, people think that oestrogenic hormon is in conjunction with intracellular single ER.As described below, when having cloned second kind of ER (ER-β) (the sort of ER originally is called as ER-α) and found to regulate the cofactor that ER replys, this simple viewpoint has been changed fully.Part can be in conjunction with two kinds of different ER, and when having tissue specificity co-activation thing and/or corepressor, these two kinds of different ER are in conjunction with the oestrogenic hormon response element in generegulation district or in conjunction with other transcription factor.Because the complicacy of ER signal and ER-α and ER-β and their cofactor tissue specific expression, the ER part can be used as estrogen agonist and antagonist so it is now know that, and it is simulated estrogenic positive effect or prevent negative effect in the tissue specificity mode.This makes us find the medicine that is called selective estrogen receptor modulators (Selective Estrogen Receptor Modulator) or SERM that a class is brand-new.These medicines have and prevent and/or treat cancer and osteoporosis significantly, and the potential of neurodegenerative diseases such as cardiovascular disorder and degenerative brain disorder.

Bone-resorbing disease as osteoporosis, be the many crowds' of influence senile symptom, and its methods of treatment is limited.For example, in the U.S., the philtrum more than 50 years old has 50% the women and Yue 10% the male sex to suffer from osteoporosis approximately.In suffering from osteoporotic individuality, the bone material runs off to increase and makes the bone fragility, and consequently risk of bone fracture increases.Other bone-resorbing disease such as Paget's disease and metastatic bone cancer, also has similar symptom.

Bone is a kind of living tissue that contains the number of different types cell.In healthy individual, the amount balance of the amount of the bone that scleroblast is made and osteoclast eliminating or resorbent bone.In suffering from the individuality of bone-resorbing disease, the function imbalance of these two types of cells.The modal example that this imbalance is possible is that postmenopausal women's bone resorption increases sharply.The bone loss of this acceleration is that the estrogen deficiency relevant with menopause causes.Yet, among the mechanism how estrogenic loss causes bone resorption to increase is being argued always.

Recently, the researchist thinks, the increase of interleukin 1 (" IL-1 ") and tumour necrosis factor bone resorption cytokines such as (" TNF ") may be the reason (Kimble etc. that bone runs off after the menopause, J.Biol.Chem.271:28890-28897,1996), and the inhibitor of these cytokines can partly reduce the bone loss (Pacific J.Bone Miner Res.11:1043-1051,1996) of rodent oophorectomize postoperative.In addition, reported once that stopping to produce oestrogenic hormon can make murine marrow and osteocyte secretion IL-6 increase (Girasole etc., J.Clin.Invest.89:883-891,1992; Jilka etc., Science257:88-91,1992; Kimble etc., Endocrinology 136:3054-3061,1995; Passseri etc., Endocrinology 133:822-828,1993), the antibody of IL-6 can suppress the increase (Girasole etc. of the depleted mouse osteoclast precursor of oestrogenic hormon, the same), and the bone that the oophorectomize postoperative can not take place in lacking the transgenic mice of IL-6 run off (Poli etc., EMBO J.13:1189-1196,1994).

The existing treatment that slows down the bone loss generally includes the compound of using oestrogenic hormon, diphosphate, thyrocalcitonin and raloxifene and so on.But these compounds are generally used for long-term treatment, and adverse side effect is arranged.In addition, this treatment can cause sophisticated osteoclast activation usually rather than reduce their formation.For example, estrogen-induced osteoclast apoptosis and thyrocalcitonin dwindles osteoclast and separate (Hughes etc., Nat.Med.2:1132-1136,1996 from bone surface; Jilka etc., Exp.Hemato.23:500-506,1995).Equally, diphosphate reduces osteoclast activity, changes its form and increases apoptosis (Parfitt etc., J.Bone MinerRes.11:150-159,1996 of osteoclast; Suzuki etc., Endocrinology 137:4685-4690,1996).

Think that also cytokine also plays an important role in various cancers.For example, the researchist shows, IL-6 is autocrine/paracrine somatomedin (Seigall etc., Cancer Res.50:7786,1999) in prostate cancer, can improve the survival rate (Okamoto etc. of tumour, Cancer Res.57:141-146,1997), and in and IL-6 antibody can reduce cell proliferation (Okamoto etc., Endocrinology138:5071-5073,1997; Borsellino etc., Proc.Annu.Meet.Am.Assoc.Cancer Res.37:A2801,1996).It is reported that IL-6 is for multiple myeloma (Martinez-Maza etc., Res.Immunol.143:764-769,1992; Kawano etc., Blood 73:517-526,1989; Zhang etc., Blood 74:11-13,1989; Garrett etc., Bone 20:515-520,1997; With Klein etc., Blood 78:1198-12-4,1991), renal cell carcinoma (Koo etc., Cancer Immunol.35:97-105,1992; Tsukamoto etc., J.Urol.148:1778-1782,1992; With Weissglas etc., Endocrinology 138:1879-1885,1997) and cervical cancer (Estuce etc., Gynecol.Oncol.50:15-19,1993; Tartour etc., Cancer Res.54:6243-6248,1994; With Iglesias etc., Am.J.Pathology 146:944-952,1995) also produce similar effects.

In addition, think that also IL-6 is relevant with sacroiliitis, especially adjuvant, collagen and antigen induced sacroiliitis (Alonzi etc., J.Exp.Med.187:146-148,1998; Ohshima etc., Proc.Natl.Acad.Sci.USA 95:8222-8226,1998; With Leisten etc., Clin.Immunol.Immunopathol56:108-115,1990), have report with resist-IL-6 antibody comes treatment of arthritis (Wendling etc., J.Rheumatol.20:259-262,1993).In addition, shown that oestrogenic hormon can induce inhibition test systemic autoimmune encephalomyelitis and collagen-induced property sacroiliitis (Jansson etc., Neuroimmunol.53:203-207,1994) in mouse.

Also showed cell factor IL-6 be a kind of important induce the factor that osteoclast forms (Girasole etc., the same; Jilka etc. (1992), the same; Jilka etc. (1995), the same; Kimble etc. (1995), the same; Pacifici etc., the same; With Passeri etc., the same).Other researchist shows that Sant 5 antagonists of using neutralizing antibody, antisense scant polymer or IL-6 have reduced number (Devlin etc., J.Bone Miner 13:393-399,1998 of osteoclast in the oophorectomize mouse spinal bone; Girasole etc., the same; Jilka etc. (1992), the same; With Schiller etc., Endocrinology138:4567-4571,1997), human cytomegalovirus absorbs ability (Ohsaki etc., Endocrinology131:2229-2234,1993 of dentine; With Reddy etc., J.Bone Min.Res.9:753-757,1994), and the formation of osteoclast in normal people's marrow culture.Find that also oestrogenic hormon reduced the activity (Stein etc., Mol.Cell Biol 15:4971-4979,1995) of IL-6 promotor by the interaction between estrogen receptor and transcription factor NF-KB and C/EBP β.

Thought that granulocyte-macrophage colony stimutaing factor (" GM-CSF ") plays a role in the propagation of broken bone precursor cell.In the extended culture of people or bone marrow cells in mice or peripheral blood cells, GM-CSF promotes osteoclast to form (Kurihara etc., Blood 74:1295-1302,1989; Lorenzo etc., J.Clin.Invest.80:160-164,1987; MacDonald etc., J.Bone Miner 1:227-233,1986; With Shinar etc., Endocrinology 126:1728-1735,1990).From the postmenopausal women or stop the women of estrin treatment isolating medullary cell and compare, express the GM-CSF (Bismar etc., J.Clin.Endocrinol.Metab.80:3351-3355,1995) of higher level with isolated cells the women before menopause.Be presented among the patient of orthopaedic implants erosion the expression of GM-CSF relevant with the distribution of bone resorption osteoclast (Al-Saffar etc., Anatomic Pathology105:628-693,1996).

As mentioned above, thought oestrogenic hormon in the past always, and caused conformational change and discharge, and the interior so-called oestrogenic hormon response element of dimer acceptor and range gene promoter region is combined from heat shock protein in conjunction with intracellular single ER.And the pharmacologist it has been generally acknowledged that non-steroid micromolecular part and ER competition conjugated estrogen hormone, in the various tissues of expressing estrogen receptor as antagonist or agonist.Therefore, only this part is divided into antagonist or agonist usually.Think that now this is not incorrect.

Think that now oestrogenic hormon is regulated cellular pharmacology by genetic expression, and estrogenic effect is mediated by estrogen receptor.As mentioned above, two kinds of estrogen receptor are arranged now, ER-α and ER-β.Estrogen receptor can mediate by following two kinds of approach the effect of generegulation: make the direct conjugated estrogen hormone oestrogenic hormon of ER response element (ERE)-" classical pathway " (Jeltsch etc., Nucleic Acids Res.15:1401-1414,1987; Bodine etc., Endocrinology 139:2048-2057,1998), make ER in conjunction with other transcription factor-" non-classical approach " (Stein etc., Mol.Cell Biol.15:4971-4979,1995 such as NF-κ B, C/EBP-β or AP-1; Paech etc., Science 277:1508-1510,1997; Duan etc., Endocrinology 139:1981-1990,1998), and can mediate (Nadal by non-genomic group effect through the outer estrogen receptor signal transduction of the nuclear that may comprise plasma membrane ER, A. etc., Trends in Pharmacological Sciences 22:597-599,2001; Wyckoff, M.H. etc., J.Biol.Chem.276:27071-27076,2001; Chung, Y-L. etc., Int.J.of Cancer97:306-312,2002; Kelly, M.J. etc., Trends Endocrinol.Metab.10:369-374,1999; Levin, E.R. etc., Trends Endocrinol.Metab.10:374-377,1999).

The progress of recent years shows that ER activator (for example SRC-I, CBP and SRA) together is relevant with corepressor (for example SMRT and N-CoR), and they also regulate the transcriptional activity of ER with tissue specificity and ligand specificity's mode.In this case, ER and the crucial transcription factor interaction of regulating these genes.Knownly can regulate active transcription factor by ER and comprise, for example AP-1, NF-κ B, C/EBP and Sp-1.In addition, identified lonely nuclear receptor, as estrogen receptor associated receptor a, b, g (ERR-a, ERR-b, ERR-g).Although it is not the part of ERR that estradiol shows, some SERM and other traditional E R part have demonstrated with high-affinity in conjunction with these acceptors (Coward, P. etc., Proc.Natl Acad.Sci.98:8880-8884,2001; Lu, D. etc., CancerRes.61:6755-6761,2001; Tremablay, G.B. etc., Endocrinology 142:4572-4575,2001; Chen, S. etc., J.Biol.Chem.276:28465-28470,2001).

In addition, owing to lack good ER-β antibody, mainly be to analyze therefore by RT-PCR or in situ hybridization, analysis shows ER-α and the existing overlapping different tissue distribution that has again of ER-β.Yet owing to be used for measuring the differentiation state of the method for ER, the species of being analyzed (rat, mouse, the mankind) and/or isolating primary cell, some of them result is controversial.Usually organize and both expressed ER-α, also express ER-β, but these receptor mappings are in different cellular type.In addition, some tissues (as kidney) only contain ER-α, and other tissue (as uterus, hypophysis and epididymis (epidymis)) in ER-α account for very big advantage (Couse etc., Endocrinology 138,4613-4621,1997; Kuiper etc., Endocrinology138,863-870,1997).On the contrary, the tissue of expressing high-level ER-β comprises some zone (Brandenberger etc., J.Clin.Endocrinol.Metab.83,1025-8,1998 of prostate gland, testis, ovary and brain; Enmark etc., J.Clinic.Endocrinol.Metabol.82,4258-4265,1997; Laflamme etc., J.Neurobiol.36,357-78,1998; Sar and Welsch, Endocrinology 140,963-71,1999; Shughrue etc., Endocrinology 138,5649-52,1997a; Shughrue etc., J.Comp.Neurol.388,507-25,1997b).

The foundation of ER-α (Korach, Science 266,1524-1527,1994) and ER-β (Krege etc., Proc.Natl.Acad.Sci.USA 95,15677-82,1998) knock-out mice has confirmed that also ER-β has different functions in different tissues.For example, ER-α knock-out mice (male and female) is sterile, female not demonstration property susceptibility and malely do not have typical male aggressiveness behavior (Cooke etc., Biol.Reprod.59,470-5,1998; Das etc., Proc.Natl.Acad.Sci.USA 94,12786-12791,1997; Korach, 1994; Ogawa etc., Proc.Natl.Acad.Sci.USA 94,1476-81,1997; Rissman etc., Endocrinology 138,507-10,1997a; Rissman etc., Horm.Behav.31,232-243,1997b).In addition, the brain of these animals is still to reply (Shughrue etc. with the similar pattern of wild-type animal to oestrogenic hormon, Proc.Natl.Acad.Sci.USA 94,11008-12,1997c), and oestrogenic hormon still suppresses because the mechanical wounding blood vessel injury (Iafrati etc., the Nature Med.3 that cause, 545-8,1997).On the contrary, the mice develop that lacks ER-β is normal, can give birth to and shows normal sexual behaviour, farrows less and less (Krege etc., 1998) but compare the every nest of wild-type mice, and mammogenesis is normal and lactation is normal.Fertility reduces the result who is considered to the reduction of ovary effect, and ER-β is an ER form main in the ovary, and it is arranged in the granulosa cell of ripe follicle.

In a word, know the medicinal use of compound in the multiple oestrogenic hormon related symptoms of treatment for a long time as estrogen antagonist or agonist, described symptom comprises the symptom (Barkhem etc. relevant with brain, bone, cardiovascular systems, skin, hair follicle, immunity system, bladder and prostate gland, Mol.Pharmacol.54,105-12,1998; Farhat etc., FASEB J.10,615-624,1996; Gustafsson, Chem.Biol.2,508-11,1998; Sun etc., 1999; Tremblay etc., Endocrinology 139,111-118,1998; Turner etc., Endocrinology 139,3712-20,1998).In addition, described multiple mammary gland and non-breast cancer cell and expressed ER, and target tissue (Brandenberger etc., 1998 of the concrete estrogen antagonist of conduct; Clinton and Hua, Crit.Rev.Oncol.Hematol.25,1-9,1997; Hata etc., Oncology 55 supplementary issues 1,35-44,1998; Rohlff etc., Prostate 37,51-9,1998; Simpson etc., J Steroid Biochem Mol Biol 64,137-45,1998; Yamashita etc., Oncology 55 academic periodicals 1,17-22,1998).

Developed many in recent years and interactional steroid class of ER and non-steroidal compounds.For example, tamoxifen is as antiestrogen at first and is used for treating mammary cancer, but recent findings it in uterus, bone and cardiovascular systems, can be used as the part estrogen agonist.Raloxifene is another kind of compound of once advising as SERM, but has confirmed to can be used to treat osteoporosis.

The tamoxifen raloxifene

Also reported the analogue (Grese etc., J.Med.Chem.40:146-167,1997) of raloxifene.

As for the tonka bean camphor based compound, mentioned many structures, comprise following these: U.S. Patent number 6,291,456,6,331,562 and 6,593,322; Roa etc., Synthesis 887-888,1981; Buu-Hoi etc., J.Org.Chem.19:1548-1552,1954; Gupta etc., Indian J.Exp.Biol.23:638-640,1985; Disclosed PCT application number WO 96/31206; Verma etc., Indian J.Chem.32B:239-243,1993; Lednicer etc., J.Med.Chem.8:725-726,1965; Micheli etc., Steroids 5:321-335,1962; Brandt etc., Int.J.QuantumChemistry:Quantum Biol.Symposia 13:155-165,1986; Wani etc., J.Med.Chem.18:982-985,1975; Pollard etc., Steroids 11:897-907,1968.

Therefore, this area needs improved compound to be used for treating or preventing the disease of bone-resorbing disease, neoplastic disease, sacroiliitis, the disease that worsens or improvement when oestrogenic hormon exists when oestrogenic hormon exists; Be used for activating the function of ER in the osteocyte, the function of ER in the anticancer suppresses the expression of IL-6 in the cell and suppresses growth of tumour cell.

Any bibliography that the application's part 2 is quoted and mentioned can not be thought the application's prior art.

3. summary of the invention

The present invention relates to have the compound and the pharmacy acceptable salt thereof of following structural formula (I):

In the formula:

X and Y are hydrogen, halogen or (halo) (C independently _1-6Alkyl);

N is 1,2 or 3; With

One of following situation:

(a) R ₁Be hydrogen, R ₂Be halogen, hydroxyl ,-(C _1-6Alkyl), vinyl ,-(C _2-6Alkynyl) ,-C (O) O (C _1-6Alkyl), (C-(hydroxyl) _1-6Alkyl), (C-(amino) _1-6Alkyl) ,-(CH ₂) _m-O-(C _1-6Alkyl) ,-C (O) (C _1-6Alkyl) ,-(CH ₂) _m-phenyl ,-(3 to 7 yuan of monocyclic heterocycles) ,-COOH ,-C (O) H or-CN;

(b) R ₂Be hydrogen, R ₁Be halogen, hydroxyl ,-(C _1-6Alkyl) ,-(C _2-6Thiazolinyl)-(C _2-6Alkynyl) ,-C (O) O (C _1-6Alkyl), (C-(hydroxyl) _1-6Alkyl), (C-(amino) _1-6Alkyl) ,-(CH ₂) _m-O-(C _1-6Alkyl) ,-C (O) (C _1-6Alkyl) ,-(CH ₂) _m-phenyl ,-(3 to 7 yuan of monocyclic heterocycles) ,-COOH ,-C (O) H or-CN; Or

(c) R ₁And R ₂Be-CH ₃

The compound of formula (I) or the pharmacy acceptable salt of this compound (all being " chromone compound ") are used to treat or prevent the disease of bone-resorbing disease, neoplastic disease, sacroiliitis, the disease that worsens or improvement when oestrogenic hormon exists when oestrogenic hormon exists; Activate the function of ER in the osteocyte; The function of ER in the anticancer; Suppress the expression of IL-6 in the cell; With the inhibition growth of tumour cell.

The invention still further relates to the chromone compound that contains significant quantity and the composition of pharmaceutically acceptable carrier or vehicle.Described composition is used to treat or prevent the disease of bone-resorbing disease, neoplastic disease, sacroiliitis, the disease that worsens or improvement when oestrogenic hormon exists when oestrogenic hormon exists; Activate the function of ER in the osteocyte; The function of ER in the anticancer; Suppress the expression of IL-6 in the cell; With the inhibition growth of tumour cell.

The invention still further relates to the method for preparing chromone compound, described method comprises makes have structural formula compound or its pharmacy acceptable salt demethylation of (II):

In the formula, R ₁, R ₂, X, Y and n be as mentioned to the definition of chromone compound.

The invention still further relates to the method for treatment or prevention bone-resorbing disease, this method comprises the chromone compound that needs the patient of this treatment or prevention significant quantity.

The invention still further relates to the method for treatment or prophylaxis of tumours disease, this method comprises the chromone compound that needs the patient of this treatment or prevention significant quantity.

The invention still further relates to treatment or prevent arthritic method, this method comprises the chromone compound that needs the patient of this treatment or prevention significant quantity.

The invention still further relates to the method for treatment or prevention disease of deterioration when oestrogenic hormon exists, this method comprises the chromone compound that needs the patient of this treatment or prevention significant quantity.

The invention still further relates to the method for treatment or prevention disease of improvement when oestrogenic hormon exists, this method comprises the chromone compound that needs the patient of this treatment or prevention significant quantity.

The invention still further relates to the method that activates the function of ER in the osteocyte, this method comprises the chromone compound that makes this osteocyte contact significant quantity.

The invention still further relates to the method for the function of ER in the anticancer, this method comprises the chromone compound that makes this cells contacting significant quantity.

The invention still further relates to the method that suppresses the expression of IL-6 in the cell, this method comprises the chromone compound of the cells contacting significant quantity that allows to express ER and IL-6.

The invention still further relates to the method that suppresses growth of tumour cell, this method comprises the chromone compound of the tumour cell contact significant quantity that allows to express ER.

The invention still further relates to treatment or prevention endometriosis; cardiovascular disorder; hypercholesterolemia; prostatomegaly; prostate cancer; fat; flush; skin effect; anxious state of mind; lose memory; menopausal syndrome; alopecia (baldness); type ii diabetes; degenerative brain disorder; the urinary incontinence; gastrointestinal tract disease; damage back vascular protection; the CNS effect; acne; cataract; hirsutism; multiple myeloma or lymphadenomatous method, this method comprise the compound as claimed in claim 1 that needs the patient of this treatment or prevention significant quantity or the pharmacy acceptable salt of this compound.

The invention still further relates to the prevention spermatogeny or with the method that is exposed to the relevant bad reproduction influence of environmental chemistry or natural hormone imbalance, this method comprises the chromone compound that needs the patient of this prevention significant quantity.

Can more fully understand the present invention by reference detailed Description Of The Invention and embodiment, embodiment is illustrating of non-limiting embodiments of the present invention.

4. detailed Description Of The Invention

4.1 definition

"-(C _1-6Alkyl) " be saturated straight chain or the side chain non-cyclic hydrocarbon that contains 1-6 carbon atom.Representational-(C _1-6Alkyl) comprising :-methyl ,-ethyl ,-n-propyl ,-normal-butyl ,-positive phenyl and-n-hexyl; And saturated branched-chain alkyl comprises :-sec.-propyl ,-sec-butyl ,-isobutyl-,-tertiary butyl ,-different phenyl, 2-methyl butyl, 3-methyl butyl, 2-aminomethyl phenyl, 3-aminomethyl phenyl, 4-aminomethyl phenyl, 2,3-dimethylbutyl etc.

"-(C _2-6Thiazolinyl) " be the straight or branched non-cyclic hydrocarbon that contains 2-6 carbon atom and contain at least one carbon-carbon double bond.Representational-(C _2-6Thiazolinyl) comprising :-vinyl ,-allyl group ,-the 1-butylene base ,-crotyl ,-isobutenyl ,-the 1-pentenyl ,-pentenyl ,-the 3-methyl-1-butene base ,-2-methyl-2-butene base ,-2,3-dimethyl-crotyl ,-the 1-hexenyl ,-the 2-hexenyl ,-the 3-hexenyl etc.Two keys of thiazolinyl can with other unsaturated group not conjugation or conjugation.

" vinyl " is-CH=CH ₂

"-(C _2-6Alkynyl) " be to contain 2-6 carbon atom and contain at least one carbon carbon triple-linked straight or branched non-cyclic hydrocarbon.Representational-(C _2-6Alkynyl) comprising :-ethynyl ,-proyl ,-the ethyl acetylene base ,-the 2-butyne base ,-the 1-pentynyl ,-the valerylene base ,-3-methyl isophthalic acid-butynyl ,-the 4-pentynyl ,-1-hexin base ,-2-hexin base ,-5-hexin base etc.The triple bond of alkynyl can with other unsaturated group not conjugation or conjugation.

" C (O) O (C _1-6Alkyl) ", wherein-(C _1-6Alkyl) as mentioned the definition, example include but not limited to :-C (O) OCH ₃,-C (O) OCH ₂CH ₃,-C (O) O (CH ₂) ₂CH ₃,-C (O) OCH (CH ₃) CH ₃Deng.

"-(CH ₂) _m-O-(C _1-6Alkyl) ", wherein-(C _1-6Alkyl) and m define as mentioned, example include but not limited to :-O-CH ₃,-CH ₂-O-CH ₃,-CH ₂-O-CH ₂CH ₃,-CH ₂-O-(CH ₂) ₂CH ₃,-(CH ₂) ₂-O-CH ₃,-(CH ₂) ₂-O-CH ₂CH ₃Deng.

" C (O) (C _1-6Alkyl) ", wherein-(C _1-6Alkyl) as mentioned the definition, example include but not limited to :-C (O) CH ₃,-C (O)-CH ₂CH ₃,-C (O)-(CH ₂) ₂CH ₃,-C (O)-(CH ₂) ₃CH ₃,-C (O)-(CH ₂) ₄CH ₃,-C (O)-(CH ₂) ₅CH ₃Deng.

"-(CH ₂) _m-phenyl ", m wherein defines as mentioned, example include but not limited to :-phenyl ,-CH ₂-phenyl ,-(CH ₂) ₂-phenyl ,-(CH ₂) ₃-phenyl ,-(CH ₂) ₄-phenyl ,-(CH ₂) ₅-phenyl etc.

" 3 to 7 yuan of monocyclic heterocycles " is to contain at least one to be selected from O, the atom of N or S also contains the monocyclic groups of 2-6 carbon atom, it can be saturated, insatiable hunger and/or fragrance, include, but is not limited to pyrrolidyl, pyrryl, pyrazolyl, oxetanyl, pyrazolinyl, imidazolyl, imidazolinyl, imidazolidyl oxazolyl oxazolidinyl isoxazoline-3-yl isoxazolyl, thiazolyl, thiadiazolyl group, thiazolidyl, isothiazolyl, the isothiazole alkyl, furyl, tetrahydrofuran base, thienyl oxadiazole base, piperidyl, piperazinyl, 2-oxo piperazinyl, 2-oxo-piperidine base, 2-oxo-pyrrolidine base, 2-oxo azepine base (2-oxazepinyl), the azepine base, the 4-piperidone base, pyridyl, N-oxo-pyridyl, pyrazinyl, pyrimidyl, pyridazinyl, THP trtrahydropyranyl, tetrahydrochysene sulfo-pyranyl, tetrahydrochysene sulfo-pyranyl sulfone, morpholinyl, thio-morpholinyl, the thio-morpholinyl sulfoxide, the thio-morpholinyl sulfone, 1,3-dioxolane and tetrahydrochysene-1,1-dioxo thienyl alkyl dioxin, the isothiazole alkyl, thietanyl, thiiranes group, triazinyl and triazolyl.

"-(amino) (C _1-6Alkyl) " by 1 or 2 amino C defined above that replaces _1-6Alkyl.Its example includes but not limited to :-CH ₂-NH ₂,-CH ₂CH ₂NH ₂,-(CH ₂) ₃NH ₂,-(CH ₂) ₄NH ₂,-(CH ₂) ₅NH ₂,-CH ₂-(NH ₂) (CH ₂) ₅CH ₃,-CH-(NH ₂) (CH ₃) ₂,-CH-(NH ₂) (CH ₂CH ₃) ₂,-C-(NH ₂) ((CH ₂) ₂CH ₃) ₂Deng.

"-(hydroxyl) (C _1-6Alkyl) " C defined above that is replaced by 1 or 2 hydroxyl _1-6Alkyl.Its example includes but not limited to :-CH ₂OH ,-(CH ₂) ₂OH ,-(CH ₂) ₃OH ,-(CH ₂) ₄OH ,-(CH ₂) ₅OH ,-CH (OH) CH ₃,-CH (OH) CH ₂CH ₃,-CH ₂CH (OH) CH ₃Deng.

" halogen " is fluorine, chlorine, bromine or iodine.

"-(halo) (C _1-6Alkyl) " C defined above that is replaced by one or more halogens _1-6Alkyl.Its example includes but not limited to :-CF ₃,-CHF ₂,-CH ₂F ,-CCl ₃,-CHCl ₂,-CBr ₃,-CHBR ₂,-CH ₂CF ₃,-CH ₂CHF ₂,-CH ₂CH ₂F etc.

Described chromone compound can have chiral centre and can be used as racemoid, independent enantiomer or diastereomer and their mixture occurs.All these isomeric forms all are included within the present invention, and comprise their mixture.In addition, some chromone compounds can be used as polymorphic form and exist, and this is also included within the present invention.In addition, some chromone compounds also can form solvate with water or other organic solvent.This solvate equally also within the scope of the invention.

Oestrogenic hormon " agonist " be a kind of in one or more cell or tissues in conjunction with ER and activate the compound of the function of ER, and oestrogenic hormon " antagonist " in one or more cell or tissues in conjunction with ER and suppress the function of ER.ER comprises the isotype of ER-α, ER-β, ER-α or ER-β or mutant and with ER the protein of at least 95% homology is arranged.

Term " significant quantity " and chromone compound coupling represent to realize the amount of following effect: treatment bone-resorbing disease, neoplastic disease, sacroiliitis, the disease that worsens when oestrogenic hormon exists or the disease of improving when oestrogenic hormon exists; Activate the function of ER in the osteocyte; The function of ER in the anticancer; Suppress the expression of IL-6 in the cell; Or inhibition growth of tumour cell.

Term " significant quantity " and other therapeutical agent coupling represent to realize the amount of following effect: treatment or prevention bone-resorbing disease, neoplastic disease, sacroiliitis, the disease that worsens when oestrogenic hormon exists or the disease of improving when oestrogenic hormon exists; Activate the function of ER in the osteocyte; The function of ER in the anticancer; Suppress the expression of IL-6 in the cell; Or the inhibition growth of tumour cell, described chromone compound is brought into play its treatment or prophylactic effect simultaneously.

" patient " is animal, includes but not limited to animals such as ox, monkey, horse, sheep, pig, chicken, turkey, quail, cat, dog, mouse, rat, rabbit and cavy, is Mammals in one embodiment, is being the people in another embodiment.

4.2 chromone compound

The present invention relates to have the chromone compound of following structural formula (I):

In the formula, R ₁, R ₂, X, Y and n define as mentioned.

In one embodiment, the R of described chromone compound ₁Be hydrogen.

In another embodiment, the R of described chromone compound ₂Be hydrogen.

In another embodiment, the R of described chromone compound ₁And R ₂All be-CH ₃

In another embodiment, the R of described chromone compound ₁And R ₂In one be-C (O)-(C _1-6And another is a hydrogen alkyl).

In another embodiment, the R of described chromone compound ₁And R ₂In one be-C (O) H that and another is a hydrogen.

In another embodiment, the R of described chromone compound ₁And R ₂In one be methyl, and another is a hydrogen.

In another embodiment, the R of described chromone compound ₁Be methyl, and R ₂Be hydrogen.

In another embodiment, the R of described chromone compound ₂Be methyl, and R ₁Be hydrogen.

In another embodiment, the R of described chromone compound ₁And R ₂In one be CN, and another is a hydrogen.

In another embodiment, the R of described chromone compound ₁And R ₂In one be-(hydroxyl) (C _1-6And another is a hydrogen alkyl).

In another embodiment, the R of described chromone compound ₁And R ₂In one be halogen, and another is a hydrogen.

In another embodiment, the R of described chromone compound ₁Be-(C _2-6And R thiazolinyl), ₂Be hydrogen.

In another embodiment, the R of described chromone compound ₂Be vinyl, and R ₁Be hydrogen.

In another embodiment, the X of described chromone compound and Y are halogens.

In another embodiment, the X of described chromone compound and Y are chlorine.

In another embodiment, the X of described chromone compound and Y are trifluoromethyls.

In another embodiment, the X of described chromone compound is a hydrogen, and Y is a trifluoromethyl.

In another embodiment, the X of described chromone compound is a trifluoromethyl, and Y is a chlorine.

In another embodiment, the X of described chromone compound is a chlorine, and Y is a trifluoromethyl.

In another embodiment, the X of described chromone compound is a hydrogen, and Y is a halogen.

In another embodiment, the R of described chromone compound ₁And R ₂In one be-C (O)-(C _1-6Alkyl), and another is a hydrogen, and X and Y be halogen, as chlorine.

In another embodiment, the R of described chromone compound ₁And R ₂In one be methyl, and another is a hydrogen, X and Y are halogens, as chlorine.

In another embodiment, the R of described chromone compound ₁And R ₂In one be methyl, and another is a hydrogen, one among X and the Y is trifluoromethyl, another is a hydrogen.

In another embodiment, the R of described chromone compound ₁And R ₂In one be-C (O)-(C _1-6Alkyl), and another is a hydrogen, and one among X and the Y is trifluoromethyl, and another is a hydrogen.

In another embodiment, the R of described chromone compound ₁And R ₂In one be CN, and another is a hydrogen, one among X and the Y is trifluoromethyl, another is a hydrogen.

In another embodiment, the R of described chromone compound ₁And R ₂In one be-CH (OH)-CH ₃, and another is a hydrogen, and one among X and the Y is trifluoromethyl, another is a hydrogen.

In another embodiment, the R of described chromone compound ₁And R ₂In one be halogen, as bromine, and another is a hydrogen, one among X and the Y is trifluoromethyl, another is a hydrogen.

In another embodiment, the R of described chromone compound ₁And R ₂In one be halogen, as iodine, and another is a hydrogen, one among X and the Y is halogen, as chlorine, another is a hydrogen.

In another embodiment, the R of described chromone compound ₁Be-CH ₂-CH=CH ₂, R ₂Be hydrogen, one among X and the Y is trifluoromethyl, and another is a hydrogen.

In another embodiment, the R of described chromone compound ₂Be vinyl, R ₁Be hydrogen, one among X and the Y is trifluoromethyl, and another is a hydrogen.

In another embodiment, the R of described chromone compound ₁And R ₂In one be-CH ₂-NH ₂, and another is a hydrogen, and one among X and the Y is halogen, as chlorine, another is a hydrogen.

In another embodiment, the n of described chromone compound is 1.

4.3 prepare the method for chromone compound

Described chromone compound can prepare according to the popular response among following scheme 1-23 and the embodiment 1-9.

Scheme 1:

The embodiment 1 that the illustrative example of top scheme 1 steps A-H sees below.

Scheme 2:

The embodiment 3 that the illustrative example of top scheme 2 steps A-G sees below.The embodiment 4 that top scheme 2 step H and the illustrative example of I see below.

Scheme 3:

The raw material of scheme 3 can prepare according to the description among the embodiment 9 hereinafter.

Scheme 4:

The raw material of scheme 4 can prepare according to the description in the scheme 2 above.

Scheme 5:

The raw material of scheme 5 can prepare according to the description among the embodiment 9 hereinafter.

Scheme 6:

The raw material of scheme 6 can prepare according to the description in the scheme 2 above.

Scheme 7:

The raw material of scheme 7 can prepare according to the description in the scheme 2 above.

Scheme 8:

The raw material of scheme 8 can prepare according to the description among the embodiment 9 hereinafter.

Scheme 9:

The raw material of scheme 9 can prepare according to the description in the scheme 2 above.

Scheme 10:

The raw material of scheme 10 can prepare according to the description among the embodiment 9 hereinafter.

Scheme 11:

The raw material of scheme 11 can prepare according to the description in the scheme 9 above.

Scheme 12:

The raw material of scheme 12 can prepare according to the description in the scheme 10 above.

Scheme 13:

The raw material of scheme 13 can prepare according to the description in the scheme 2 above.

Scheme 14:

The raw material of scheme 14 can prepare according to the description among the embodiment 9 hereinafter.

Scheme 15:

The raw material of scheme 15 can prepare according to the description in the scheme 2 above.

Scheme 16:

The raw material of scheme 16 can obtain by the step H that carries out such scheme 2 with the material by the hereinafter description acquisition of embodiment 9.Under 0 ℃, use NaBH ₄Handle the THF/MeOH solution of corresponding 6-ethanoyl-7-hydroxychromen-2-ketone.After reaction is finished rough reaction mixture is carried out purifies and separates and go out required product.

Scheme 17:

The raw material of scheme 17 can prepare according to the description among the embodiment 9 hereinafter.

Scheme 18:

The raw material of scheme 18 can prepare according to such scheme 15.Under 0 ℃, use NaBH ₄Handle the THF/MeOH solution of corresponding 8-ethanoyl-7-hydroxychromen-2-ketone.After reaction is finished rough reaction mixture is carried out purifies and separates and go out required product.

Scheme 19:

The raw material of scheme 19 can prepare according to the description in the scheme 2 above.

Scheme 20:

The raw material of scheme 20 can prepare according to the description among the embodiment 9 hereinafter.

Scheme 21:

The raw material of scheme 21 can prepare according to the description in the scheme 1 above.

Scheme 22:

The raw material of scheme 22 can prepare according to the description in the scheme 5 above.

Scheme 23:

The raw material of scheme 23 can prepare according to the description in the scheme 6 above.

In one embodiment, the present invention relates to prepare the method for chromone compound, comprise compound or its pharmacy acceptable salt demethylation structural formula (II):

The demethylation of the compound of formula (II) can be used for making any method of phenol methyl ester demethylation to realize with known in the art.The example of this method can be at Greene, T.W., and Protective Groups inOrganic Synthesis, 88-92 finds in (1981), and the document is included this paper in as a reference in full.In one embodiment, demethylation carries out in order to following method, this method comprises the demethylation reagent of the about 1.0-50.0 molar equivalent of compound contact that makes formula (II), as iodo trimethyl silane, pyridine hydrochloride, Hydrogen bromide (" HBr "), hydrochloric acid, hydroiodic acid HI, BBr ₃, Grignard reagent (for example RMgX, wherein R is CH ₃, C ₂H ₅, C ₆H ₅Deng hydrocarbyl group; X is halogen atoms such as chlorine, bromine or iodine), Lewis acid is (as AlBt ₃, BF ₃, BCl ₃, B (CH) ₃, Zn (OH) ₂, AgCl etc.) or nucleophilic reagent such as sulfur alcohol.In one embodiment, described demethylation reagent is the HBr aqueous solution, and it can use when having acetate.In another embodiment, demethylation is by having demethylation reagent and randomly existing under the solvent situation of (comprising carboxylic acid), realize at about room temperature to the compound of about 200 ℃ temperature underfeed furnace (II) or its pharmacy acceptable salt, in one embodiment, be to about 160 ℃ temperature, to heat 15 minutes to about 24 hours at about 100 ℃.In one embodiment, the demethylation reaction vessel seals, and for example is sealed tube, to prevent solvent evaporation, especially when the boiling point of solvent is lower than the demethylation temperature of reaction.Directly separated salt can obtain the acid salt of chromone compound from the demethylation reactant, and demethylation reagent wherein is, for example Hydrogen bromide, hydrochloric acid or hydroiodic acid HI.Wash this acid salt and the separated free alkali cpd can obtain free alkali form with the appropriate base of sodium hydroxide and so on.

Described chromone compound can exist with the form of pharmacy acceptable salt.The free alkali form that usable acid is handled chromone compound forms pharmaceutically acceptable acid salt.Appropriate organic comprises toxilic acid, fumaric acid, phenylformic acid, xitix, succsinic acid, methanesulfonic, Phenylsulfonic acid, toluenesulphonic acids, acetate, oxalic acid, trifluoroacetic acid, propionic acid, tartrate, Whitfield's ointment, citric acid, glyconic acid, lactic acid, amygdalic acid, styracin, aspartic acid, stearic acid, palmitinic acid, formic acid, oxyacetic acid, L-glutamic acid and Phenylsulfonic acid.Suitable mineral acid comprises hydrochloric acid, Hydrogen bromide, sulfuric acid, phosphoric acid and nitric acid.Additive salt comprises muriate, bromide, iodide, hydrosulfate, acid phosphate, Yi Yansuan salt, lactic acid salt, acid Citrate trianion, oleate, tannate, pantothenate, acid tartrate, gentisate, gluconate, saccharic acid salt, carbonate, ethane sulfonate, tosilate and embonate (promptly 1,1 '-methylene radical-two-(2-hydroxyl-3-naphthoate)).Term " pharmacy acceptable salt " comprises any and all acceptable salt forms.

Pharmacy acceptable salt can form with conventional known technology, for example makes chromone compound and above-mentioned suitable acid-respons.Normally high yield forms this salt under mild temperature, and only needs usually in final synthesis step that separating compound can prepare from suitable acid wash liquid.The salifiable acid of shape can be dissolved in appropriate organic solvent or aqueous organic solvent, in alkanol, ketone or ester.On the other hand, obtain the chromone compound of free alkali form if desired, can be according to known technique from the final washing step of alkalescence with its separation.For example, a kind of typical technology for preparing hydrochloride is that free alkali is dissolved in suitable solvent, and thorough drying solution (as passing through molecular sieve), blasts hydrogen chloride gas then therein.

4.4 application method

In one embodiment, the present invention relates to treat or prevent the method for bone-resorbing disease, comprise the chromone compound that needs the patient of this treatment or prevention significant quantity.Although do not wish to be subjected to following one theory, especially in the bone-resorbing disease part, we think that described chromone compound produces by the inhibitory cell factor and/or suppresses osteoclast and form and bring into play its function.Available described chromone compound is treated or the concrete bone-resorbing disease that prevents includes but not limited to: the osteolytic lesion that osteoporosis, metastatic bone cancer, hypercalcemia, orthopaedic implants cause, Paget's disease and the bone relevant with hyperparathyroidism run off.

In another embodiment, the present invention relates to treat or the method for prophylaxis of tumours disease, comprise the chromone compound that needs the patient of this treatment or prevention significant quantity.In one embodiment, described neoplastic disease is a cancer.In one embodiment, described cancer is head and neck, eye, skin, mouth, larynx, esophagus, chest, bone, blood, lung, colon, sigmoid colon, rectum, stomach, prostate gland, mammary gland, ovary, uterus, kidney, liver, pancreas, brain, intestines, heart or adrenal cancer.In another embodiment, described cancer is carcinoma of endometrium, multiple myeloma, renal cell carcinoma or cervical cancer.Described chromone compound especially can be used for treating mammary cancer, uterus carcinoma and ovarian cancer.

Described chromone compound can be used for treating or other cancer of preventing includes but not limited to: solid tumor, sarcoma, cancer, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendothelial sarcoma, synovioma, mesothelioma, ewing's tumor, leiomyosarcoma, rhabdosarcoma, colorectal carcinoma, carcinoma of the pancreas, prostate cancer, squamous cell carcinoma, rodent cancer, gland cancer, syringocarcinoma, sebaceous carcinoma, papillary carcinoma, adenocarcinoma of nipple, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, liver cancer, cholangiocarcinoma, choriocarcinoma, spermocytoma, embryonal carcinoma, the nephroblastoma, cervical cancer, tumor of testis, lung cancer, small cell lung cancer, bladder cancer, epithelial cancer, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, Kaposi sarcoma, pinealoma, hemangioblastoma, acoustic tumor, oligodendroglioma, menangioma, melanoma, neuroblastoma, retinoblastoma, neoplastic hematologic disorder, acute lymphoblastic leukemia, the acute lymphoblastic B cell leukemia, acute lymphoblastic T chronic myeloid leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute monocytic leukemia, acute erythrocytic leukemia, acute megakaryoblastic leukemia, acute myelogenous monocytic leukemia, acute nonlymphocytic leukemia, acute nondifferentiated leukemia, chronic myelocytic leukemia, lymphocytic leukemia, hairy cell leukemia or multiple myeloma.

In another embodiment, the present invention relates to treatment or prevent arthritic method, comprise the chromone compound that needs the patient of this treatment or prevention significant quantity.In one embodiment, described sacroiliitis is adjuvant, collagen, bacterium or antigen induced sacroiliitis.In another embodiment, described sacroiliitis is rheumatic arthritis.

In another embodiment, the present invention relates to treat or prevent the method for the disease of deterioration when oestrogenic hormon exists, comprise the chromone compound that needs the patient of this treatment or prevention significant quantity.In one embodiment, the described disease that worsens when oestrogenic hormon exists is mammary cancer, ovarian cancer or uterus carcinoma.In one embodiment, described oestrogenic hormon is endogenous.In another embodiment, described oestrogenic hormon is external source.

In another embodiment, the present invention relates to treat or prevent the method for the disease of improvement when oestrogenic hormon exists, comprise the chromone compound that needs the patient of this treatment or prevention significant quantity.In one embodiment, described patient's the disease of improving when oestrogenic hormon exists is a bone-resorbing disease.In one embodiment, described patient's the disease of improving when oestrogenic hormon exists is that osteolytic lesion, Paget's disease and the bone relevant with hyperparathyroidism that osteoporosis, metastatic bone cancer, hypercalcemia, orthopaedic implants cause runs off.In one embodiment, described oestrogenic hormon is endogenous.In another embodiment, described oestrogenic hormon is external source.

In another embodiment; the present invention relates to treatment or prevention endometriosis; hypercholesterolemia; prostatomegaly; prostate cancer; fat; flush; skin effect is (as xerosis cutis; produce wrinkle; attenuation or follow the string); anxious state of mind; lose memory; menopausal syndrome; alopecia (baldness); type ii diabetes; degenerative brain disorder; the urinary incontinence; gastrointestinal tract disease (as Crohn's disease or irritable bowel syndrome); damage back vascular protection; CNS effect (as flush or memory loss); acne or cataractous method comprise the chromone compound that needs the patient of this treatment or prevention significant quantity.Be not limited to any concrete theory, the applicant thinks that above-mentioned symptom improves when oestrogenic hormon exists.

In another embodiment, the present invention relates to treatment or preventing cardiovascular disease, hirsutism, multiple myeloma or lymphadenomatous method, comprise the chromone compound that needs the patient of this treatment or prevention significant quantity.Be not limited to any concrete theory, the applicant thinks that above-mentioned symptom worsens when oestrogenic hormon exists.

In another embodiment, the present invention relates to prevent spermatogenetic method, comprise the chromone compound that needs the patient of this prevention significant quantity.

In another embodiment, the present invention relates to prevent the method for the bad reproduction influence (as inborn defect, miscarriage or spontaneous abortion) relevant, comprise the chromone compound that needs the patient of this prevention significant quantity with being exposed to the imbalance of environmental chemistry or natural hormone.

In another embodiment, described chromone compound is used to oral contraception; Prevent threatened abortion or habitual abortion; Alleviate dysmenorrhoea; Alleviate anovulatory dysfunctional uterine hemorrhage; Alleviate endometriosis; Help the development of ovary; Reduce women's chaeta hypertrophy (hirsutism); Prevention or treatment atherosclerosis; And inhibition postpartum milk secretion.Described chromone compound also has beneficial effect to the blood plasma lipide level, therefore can be used for treatment and prevention hypercholesterolemia.

In another embodiment, the present invention relates to activate the method for the function of ER in the osteocyte, comprise the chromone compound of the osteocyte contact significant quantity that allows to express ER.The function that activates ER in the osteocyte can be used to treatment or preventing osteoporosis.

In another embodiment, the present invention relates to the method for the function of ER in the anticancer, comprise the chromone compound of the cancer cells contact significant quantity that allows to express ER.In one embodiment, described cancer cells is breast cancer cell, ovarian cancer cell, endometrial carcinoma cell, uterus carcinoma cell, prostate cancer cell or hypothalamus cancer cells.The function of ER can be used to cell growth inhibiting in the anticancer, therefore can be used for treatment or preventing cancer.In one embodiment, described breast cancer cell is MCF-7.In one embodiment, described ovarian cancer cell is BG-1.

In another embodiment, the present invention relates to suppress the method for the expression of IL-6 in the cell, comprise the chromone compound of the cells contacting significant quantity that allows to express ER and IL-6.In one embodiment, the cell of described expression ER and I1-6 is an osteocyte.In another embodiment, the cell of described expression ER and I1-6 is the people U-2OS osteosarcoma cell of personnel selection ER-β stable transfection.The expression that suppresses IL-6 in the cell in vivo can be used to treat bone loss disease or osteocarcinoma.In one embodiment, described bone loss disease is an osteoporosis.The expression of IL-6 can be used for bioactivity screening test (for example as standard) and suppresses the chromone compound that IL-6 expresses with screening in the vitro inhibition cell.

In another embodiment, the present invention relates to suppress the method for growth of tumour cell, comprise the chromone compound of the tumour cell contact significant quantity that allows to express ER.In one embodiment, the tumour cell of described expression ER is a cancer cells.Can express the cancer cells of ER or the example of tumour cell includes but not limited to: mammary gland cell, gonad cell, endometrial cell, uterine cell, prostatic cell and hypothalamus cells.Suppress described cancer cells or tumor cell proliferation in vivo and can be used to treatment or preventing cancer.Can be used for being used for screening the bioactivity screening test (for example as standard) of anticarcinogen or antitumour drug or being used for diagnostic assay at described cancer cells of vitro inhibition or tumor cell proliferation.

4.5 other therapeutical agent

In some embodiment, method of the present invention also comprises other other therapeutical agent that gives significant quantity.The example of other therapeutical agent includes but not limited to be used for the preparation of following purposes: treatment or prevention bone-resorbing disease, neoplastic disease, sacroiliitis, the disease that worsens when oestrogenic hormon exists or the disease of improving when oestrogenic hormon exists; Activate the function of ER in the osteocyte; The function of ER in the anticancer; Suppress the expression of IL-6 in the cell; With the inhibition growth of tumour cell.Described other therapeutical agent can be before chromone compound, afterwards or with use simultaneously.In some embodiments, chromone compound is brought into play the time of curative effect and other therapeutical agent is brought into play curative effect to the patient time-interleaving to the patient.

In other embodiments, described other therapeutical agent is used to the disease for the treatment of or preventing to improve when oestrogenic hormon exists.Other therapeutical agent of the disease that is used for treating or prevents to improve when oestrogenic hormon exists includes but not limited to: tamoxifen, raloxifene, medroxyprogesterone, Metronidazole (danizol) and gestrinone.

In one embodiment, described other therapeutical agent is used to treatment or prevention bone loss disease (for example osteoporosis).Be used for treating or prevent other therapeutical agent of bone loss disease to include but not limited to: cathepsin K inhibitor (for example propetide of cathepsin K), (for example Chinese mugwort is held in the palm phosphonate (eitodronate) to diphosphate, pamldronate, alendronate, risedronate, zolendronate salt (zolendronate), ibandronate, clodronate or Tiludronate), parathyroid hormone (parathryoid hormone) (" PTH ") or its fragment, discharge compound (for example PTH liberin) and thyrocalcitonin or its fragment of endogenous PTH.

In another embodiment, described other therapeutical agent is used to treatment or prophylaxis of tumours disease.In one embodiment, described other therapeutical agent is used to treatment or preventing cancer (for example mammary gland, ovary, uterus, prostate gland or hypothalamic cancer).Be used for treating or other therapeutical agent of preventing cancer or neoplastic disease includes but not limited to: alkylating agent (for example nitrosourea), metabolic antagonist (for example methotrexate or hydroxyurea), Etoposide, campathecins, bleomycin, Dx, daunorubicin, colchicine, irinotecan, camptothecine, endoxan, 5 FU 5 fluorouracil, cis-platinum, carboplatin, methotrexate, trimetrexate, Erbitux (Erbitux), Thalidomide, taxol, vinca alkaloids (for example vinealeucoblastine(VLB) or vincristine(VCR)) or microtubule stabilizer (for example ebormycine).

Be used for treating or other illustrative example of the therapeutical agent of preventing cancer includes but not limited to: U 42126, aclarubicin, the hydrochloric acid acodazole, acronine, U 73975, rIL-2, altretamine, Duazomycin C, the acetic acid ametantrone, aminoglutethimide, amsacrine, Anastrozole, Antramycin, Asparaginase, asperline, azacitidine, Azatepa, Azotomycin, Batimastat, benzodepa, bicalutamide, the hydrochloric acid bicalutamide, the methylsulfonic acid Bisnafide, U 77779, bleomycin sulfate, brequinar sodium, Bropirimine, busulfan, sanarnycin, calusterone, caracemide, carbetimer, carboplatin, carmustine, carubicin hydrochloride, U 80244, Cedefingol, Chlorambucil, U 12241, cis-platinum, CldAdo, the methylsulfonic acid crisnatol, endoxan, cytosine arabinoside, Dacarbazine, dactinomycin, daunorubicin hydrochloride, Decitabine, U 78938, Dezaguanine, the methylsulfonic acid Dezaguanine, diaziquone, docetaxel, Zorubicin, doxorubicin hydrochloride, droloxifene, K-21060E, dromostanolone propionate, duazomycin, edatrexate, Vaniqa, elsamitrucin, enloplatin, enpromate, Epipropidine, epirubicin hydrochloride, R 55104, esorubicin hydrochloride, estramustine, estramustine phosphate sodium, etanidazole, Etoposide, the phosphoric acid Etoposide, Etoprine, CGS-16949A, fazarabine, fenretinide, azauridine, fludarabine phosphate, Fluracil, flurocitabine, Fosquidone, Phosphotrienin sodium, gemcitabine, gemcitabine hydrochloride, hydroxyurea, idarubicin hydrochloride, ifosfamide, Thio ALP, ImiDs, interleukin-II (comprise recombinant interleukin II, or rIL2), IF2 a, Interferon Alpha-2b, Interferon, rabbit a-nl, Interferon, rabbit a-n3, interferon beta-Ia, interferon-gamma-Ib, iproplatin, U 101440E, lanreotide acetate, letrozole, leuprorelin acetate, liarozole hydrochloride, lometrexol sodium, lomustine, losoxantrone hydrochloride, masoprocol, maytenin, mustine hydrochlcride, Magace, melengestrol acetate, melphalan, menogaril, mercaptopurine, methotrexate, methotrexate sodium, U-197, meturedepa, Mitindomide, Mi Tekaxin, mitochromine, mitogillin, Mitomalcin, mitomycin, mitosper, mitotane, mitoxantrone hydrochloride, Mycophenolic Acid, R 17934, nogalamycin, ormaplatin, oxisuran, taxol, pegaspargase, peliomycin, neptamustine, Peplomycin Sulfate, perfosfamide, pipobroman, piposulfan, the hydrochloric acid piroxantrone, Plicamycin, plomestane, porfimer sodium, porfiromycin, prednimustine, procarbazine hydrochloride, puromycin, puromycin hydrochloride, pyrazofurin, riboprine, Rogletimide, Safingol, the hydrochloric acid Safingol, SelCid, semustine, simtrazene, Sparfosate Sodium, sparsomycin, spirogermanium hydrochloride, spiromustine, spiral shell platinum, Streptonigrin, streptozocin, sulofenur, talisomycin, tecogalan sodium, Tegafur, teloxandrone hydrochloride, temoporfin, teniposide, teroxirone, Testolactone, tiamiprine, Tioguanine, Temozolomide, temodar, plug is for group, sulphur azoles symbol quinoline, Win-59075, FC-1157a, trestolone acetate, the phosphoric acid triciribine, trimetrexate, the glucuronic acid trimetrexate, triptorelin, tubulozole hydrochloride, Uramustine, uredepa, vapreotide, Visudyne, Vinblastine sulphate, vincristine sulphate, vindesine, vindesine sulfate, the sulfuric acid vinepidine, the sulfuric acid vinglycinate, the sulfuric acid vinleurosine, vinorelbine tartrate, the sulfuric acid vinrosidine, the sulfuric acid vinzolidine, vorozole, zeniplatin, zinostatin, zorubicin hydrochloride.

Be used for treating or other therapeutical agent of preventing cancer includes but not limited to: 20-table-1; the 25-dihydroxy vitamin d3; 5-ethinyluracil; Abiraterone; aclarubicin; the acyl group fulvene; adecypenol; U 73975; rIL-2; the ALL-TK antagonist; altretamine; ambamustine; amidox; amifostine; aminol evulinic acid; amrubicin; amsacrine; anagrelide; Anastrozole; rographolide; angiogenesis inhibitor; antagonist D; antagonist G; Antarelix; anti--dorsal part morphogenetic proteins-1; antiandrogen; the prostatitis adenosquamous carcinoma; antiestrogen; antitumour drug; the aphidicolin glycinate; the apoptogene conditioning agent; the apoptosis instrumentality; apurinic acid; ara-CDP-DL-PTBA; the arginine deaminase; asulacrine; Atamestane; atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron; the diazonium toxin; azepine tyrosine; baccatin III derivative; balanol; Batimastat; the BCR/ABL antagonist; the benzo chlorin; the benzoyl Staurosporine; the beta-lactam derivative; β-alethine; betaclamycin B; betulinic acid; the bFGF inhibitor; bicalutamide; bisantrene (bisantrene); bisaziridinylspermine; Bisnafide; bistratene A; U 77779; breflate; Bropirimine; budotitane; buthionine sulphoximine; calcipotriol; press down the plain C of kinases; camptothecin derivative; canary pox IL-2; capecitabine; carboxamide-amino-triazole; NSC 609974; CaRestM3; CARN 700; cartilage deutero-inhibitor; U 80244; casein kinase 2 enzyme inhibitors (ICOS); cell cycle inhibitor (for example; flavones pyrrole alcohol A; tryprostatin B; pl9ink4D); cell cycle protein dependent kinase inhibitor (for example; roscovitine; olomucine and purine analogue); map kinase inhibitor (CNI-1493); castanospermine; cecropin B; cetrorelix; chlorlns; chloro quinoxaline sulphonamide (chloroquinoxalinesulfonamide); cicaprost; the cis porphyrin; CldAdo; the Clomiphene analogue; clotrimazole; collismycin A; collismycin B; combretastatin A4; the combretastatin analogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8; cryptophycin A derivative; curacinA; encircle penta anthraquinone; cycloplatam; cypemycin; arabinosylcytosine; the cytolysis factor; cytostatin; Dacliximab; Decitabine; the dehydrogenation didemnin B; deslorelin; dexamethasone; right ifosfamide (dexifosfamide); dexrazoxane; dexverapamil; diaziquone; didemnin B; didox; diethyl removes the first spermine; dihydro-U-18496; the dihydro taxol; 9-dioxolamycin; the phenylbenzene spiromustine; docetaxel; behenyl alcohol; dolasetron; doxifluridine; droloxifene; dronabinol; duocarmycin SA; ebselen; Ecomustine; Ro 14-5243; Edrecolomab; eflornithine; olive alkene; emitefur; epirubicin; epristeride; the estramustine analogue; estrogen agonist; estrogen antagonist; etanidazole; the phosphoric acid Etoposide; Exemestane; fadrozole; fazarabine; fenretinide; filgrastim; finasteride; flavones pyrrole alcohol; fluorine thiophene department spit of fland; the fluorine sterone; fludarabine; fluorodaunorunicin hydrochloride; forfenimex; formestane; Phosphotrienin; fotemustine; gadolinium moral porphyrin (gadolinium texaphyrin); gallium nitrate; Galocitabine; Ganirelix; the gelatinase inhibitor; gemcitabine; the gsh inhibitor; hepsulfam; transfer albumen; hexamethylene bisacetamide; hypericin; Ibandronic acid; idarubicin; idoxifene; Idramantone; Thio ALP; Ilomastat; the imidazoles dihydroketoacridine; Imiquimod; mucin peptide; the IGF-1R inhibitor; the Interferon, rabbit agonist; Interferon, rabbit; interleukin-; m-iodobenzylguanidine; iododoxorubicin; Mexician scammony; 4-; Iroplact; irsogladine; isobengazole; isohomohalicondrin B; U 98079A; jasplakinolide; kahalalide F; sheet spiral shell element-N triacetate; Lanreotide; leinamycin; lenograstim; the sulfuric acid lentinan; leptolstatin; letrozole; leukaemia inhibitory factor; the leukemia interferon-alpha; Leuprolide+oestrogenic hormon+Progesterone; Leuprolide; LEVAMISOLE HCL; liarozole; linear polyamine analogue; lipotropy two glycopeptides; the lipotropy platinic compound; lissoclinamide7; Lip river platinum; lombricine; lometrexol; lonidamine; losoxantrone; lovastatin; Loxoribine; lurtotecan; lutetium moral porphyrin; lysofylline; the cytolysis peptide; maitansine; seminose statin A; Marimastat; masoprocol; the arteries and veins silk is flat; bamboo peach lysin inhibitor; matrix metallo-proteinase inhibitor; menogaril; Mei Balong (merbarone); meterelin; egg ammonia enzyme (Methioninase); metoclopramide; the MIF inhibitor; mifepristone; miltefosine; mirimostim; the double-stranded RNA of mispairing; mitoguazone; mitolactol; mitomycin analogs; Mitonafide; mitomycin fibroblast growth factor-saporin; mitoxantrone; Ro 40-8757; Sch-39300; monoclonal antibody; physex; monophosphoryl lipid A+ mycobacterium cell walls sk; mopidamol; multiple medicine drug resistant gene inhibitor; treatment based on kinds of tumors inhibition 1; mustard anticancer agent; Indian Ocean sponge (mycaperoxide) B; the mycobacterium cell wall extracts; myriaporone; the N-Tacedinaline; the benzamides that N-replaces; nafarelin; nagrestip; naloxone+pentazocine; napavin; naphterpin; Neu-up 100; S 254; Nemorubicin; neridronic acid; neutral endopeptidase; Nilutamide; the silt mycin; the nitrogen oxide conditioning agent; the nitroxide antioxidant; nitrullyn; O6-benzyl guanine; Sostatin; okicenone; oligonucleotide; onapristone; ondansetron; ondansetron; oracin; oral cytokine induction agent; ormaplatin; osaterone; oxaliplatin; oxaunomycin; taxol; paclitaxel analogs; D51-7059; send labor amine; palmitoylrhizoxin; Pamidronic Acid; panoxatriol; panomifene; parabactin; Pazelliptine; pegaspargase; peldesine; Pentosan Polysulfate Sodium; pentostatin; pentrozole; Perflubron; perfosfamide; sinapyl alcohol (perillylalcohol); the azophenlyene mycin; phenylacetate; inhibitors of phosphatases; molten chain bacterium; Pilovisc; pirarubicin; piritrexim; placetin A; placetin B; Profibrinolysin activator inhibitor; platinum complexes; platinic compound; platinum-three amine compound; porfimer sodium; porfiromycin; prednisone; propyl group two-dihydroketoacridine; prostaglandin(PG) J2; proteasome inhibitor; based on the proteic immunomodulator of A; inhibitors of protein kinase C; inhibitors of protein kinase C; little algae; tyrosine phosphatase inhibitors; the purine nucleoside phosphatase inhibitor; purpurin; pyrazoloacridine; the oxyphorase polyoxyethylene conjugate of trembling of pyrrole; the raf antagonist; Raltitrexed; ranimustine; vitamin A acid (for example 9-cis RA); histone deacetylase inhibitor (Sodium propanecarboxylate for example; suberin aniline hydroxamic acid (suberoylanilide hydroxamicacid)); TRAIL; the ras farnesyl protein transferase inhibitors; the ras inhibitor; the ras-GAP inhibitor; the retelliptine of demethylation; rhenium Re 186 etidronates; rhizomycin; ribozyme; RII looks yellow acid amides; Rogletimide; rohitukine; romurtide; Roquinimex; rubiginone B1; ruboxyl; Safingol; saintopin; SarCNU; sarcophytol A; Sargramostim; the Sdi1 stand-in; semustine; astogeny deutero-inhibitor 1; the sense organ oligonucleotide; signal transduction inhibitor; signal transduction modulators; single chain antigen binding protein; sizofiran; sobuzoxane; sodium borocaptate; sodium phenylacetate; solverol; SM-binding protein; sonermin; sparfosic acid; spicamycin D; spiromustine; Si Naipanding; sponge element (spongistatin) 1; shark amine; stem cell inhibitors; the differentiation of stem cells inhibitor; stipiamide; the stromelysin inhibitor; sulfinosine; superactivity vasoactive intestinal peptide antagonist; suradista; Suramine; pisatin; the synthetic glycosaminoglycan; tallimustine; methiodide acid tamoxifen; tauromustine; Tazarotene; tecogalan sodium; Tegafur; tellurapyrylium; the Telomere terminal transferase inhibitor; temoporfin; Temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine; thaliblastine; thiocoraline; the thrombosis element; the thrombosis mimetics; Thymosin-Alpha1; the thymopoietin receptor stimulant; Thymotrinan; Triiodothyronine; ethyl group etioporphyrin tin (tin ethyl etiopurpurin); Win-59075; titanocene dichloride; topsentin; toremifene; the myeloid-lymphoid stem cell factor; translational inhibitor; tretinoin; triacetyluridine; triciribine; trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinase inhibitor; tyrphostin (tyrphostins); the UBC inhibitor; ubenimex; the urogenital sinus growth inhibiting factor of deriving; the urokinase receptor antagonist; vapreotide; variolinB; carrier system, the red corpuscle gene therapy; velaresol; veratramin(e); verdins; Visudyne; vinorelbine; vinxaltine; vitaxin; vorozole; Zanoterone; zeniplatin; zilascorb and Zinostatin stimalamer.Preferred other anticarcinogen is 5 FU 5 fluorouracil and formyl tetrahydrofolic acid.

4.6 pharmaceutical composition and route of administration

Described chromone compound can give the patient by oral or parenteral by conventional dosage form, and described dosage form is capsule, microcapsule, tablet, particle, powder, lozenge, pill, suppository, injection, suspension agent or syrup for example.The organic or inorganic additive of available routine prepares suitable formulation by common method, this additive such as vehicle (sucrose for example, starch, N.F,USP MANNITOL, sorbyl alcohol, lactose, glucose, Mierocrystalline cellulose, talcum, calcium phosphate or lime carbonate), tackiness agent (Mierocrystalline cellulose for example, methylcellulose gum, hydroxy-methyl cellulose, polypropylpyrrolidone, Polyvinylpyrolidone (PVP), gelatin, gum arabic, polyoxyethylene glycol, sucrose or starch), disintegrating agent (starch for example, carboxymethyl cellulose, hydroxypropylated starch, the low hydroxypropylcellulose that replaces, sodium bicarbonate, calcium phosphate or citrate of lime), lubricant (Magnesium Stearate for example, light anhydrous silicic acid, talcum or sodium lauryl sulphate), seasonings (citric acid for example, menthol, glycine or orange powder), sanitas (Sodium Benzoate for example, sodium bisulfite, methyl p-hydroxybenzoate or propylparaben), stablizer (citric acid for example, Trisodium Citrate or acetate), suspension agent (methylcellulose gum for example, polyvinylpyrrolidone or aluminum stearate), dispersion agent (for example Vltra tears), thinner (for example water) and basic wax (theobroma oil for example, white vaseline or polyoxyethylene glycol).The significant quantity of chromone compound can produce the level of required effect for meeting in the described pharmaceutical composition, for example, can be about 0.005-10 mg/kg weight in patients in the unitary dose for oral and administered parenterally.

Above-mentioned chromone compound usually can be with unitary dose administration every day of about 0.005-10 mg/kg weight in patients 1-4 time, but above-mentioned dosage can carry out appropriate change according to patient's age, body weight and medical conditions and administering mode.In one embodiment, this dosage is about 0.01-5 mg/kg weight in patients, about 0.05-1 mg/kg weight in patients, about 0.1-0.75 mg/kg weight in patients, or about 0.25-0.5 mg/kg weight in patients.In one embodiment, give a kind of dosage every day.

The dosage that gives patient's chromone compound can have very big variation, and this depends on the judgement of healthcare practitioner.The scope of effective administration ratio of chromone compound is generally about 0.01-5 mg/kg weight in patients, about 0.05-1 mg/kg weight in patients, about 0.1-0.75 mg/kg weight in patients, or about 0.25-0.5 mg/kg weight in patients.Certainly, usually the different constantly gradation of per daily dose in one day of chromone compound are given in practice.Yet under any particular case, the dosage of chromone compound will depend on factors such as stability such as active compound, used formulation and route of administration.

Consider that for convenience chromone compound can pass through oral administration.Yet, by in intradermal, intramuscular, intraperitoneal, transdermal, intravenously, subcutaneous, the nose, in the epidural, hypogloeeis, brain, intravaginal, through skin, rectum, suction, or topical administration ear, nose, eye or skin to give this chromone compound effective too.Preferred mode of administration depends on the judgement of healthcare practitioner, and depends in part on medical conditions.

The invention still further relates to the chromone compound that contains significant quantity and the composition of pharmaceutically acceptable carrier or vehicle, wherein said pharmaceutically acceptable carrier or vehicle can comprise vehicle, thinner or its mixing.In one embodiment, described composition is a pharmaceutical composition.

The form of described composition can be tablet, the tablet that can chew, capsule, solution, injection solution, lozenge, suppository and suspension agent etc.Composition can be made into the form that dose unit is the part of per daily dose or per daily dose, and described dose unit can be a slice tablet or the capsule or the liquid of proper volume.In one embodiment, solution prepares from water-soluble salts such as hydrochlorides.Usually, all compositions all are to prepare with the known method in pharmaceutical chemistry field.Can be with chromone compound and suitable carriers or mixing diluents, an amount of mixture is filled into prepares capsule in the capsule then.Carrier and thinner commonly used include but not limited to: the inert powder material, as various starch; Comminuted fibres element, especially crystal or Microcrystalline Cellulose; Carbohydrate is as fructose, N.F,USP MANNITOL and sucrose; Cereal powder and similar edible powder.

Can pass through wet granulation or the direct compressed tablets of non-slurry pelletizing.Usually contain thinner, tackiness agent, lubricant and disintegrating agent and compound in their prescription.Typical thinner comprises, the carbohydrate of inorganic salt such as for example various types of starch, lactose, N.F,USP MANNITOL, kaolin, calcium phosphate or calcium sulfate, sodium-chlor and pulverizing.The plain derivative of also available comminuted fibres.Typical tablet binder has the material such as carbohydrates such as starch, gelatin and lactose, fructose, glucose etc.Also commonly used natural and synthetic is gummy, comprising gum arabic, alginate, methylcellulose gum, polyvinylpyrrolidone/ etc.Polyoxyethylene glycol, ethyl cellulose and wax also can be used as tackiness agent.

In tablet, may need lubricant to be bonded on the mould to prevent tablet and drift.Lubricant can be selected from talcum, Magnesium Stearate and calcium stearate, stearic acid and the smooth solid of hydrogenant plant wet goods.Tablet disintegrant is to expand when wetting and make tablet rupture and discharge the material of compound.They comprise starch, clay, Mierocrystalline cellulose, phycocolloid and natural gum.More specifically, can use natural sponge, Zeo-karb, alginic acid, guar gum, citrus pulp and the carboxymethyl cellulose of for example corn and yam starch, methylcellulose gum, agar, wilkinite, lignocellulose, pulverizing, and sodium lauryl sulphate.Tablet can carry out dressing as seasonings and sealing agent with sugar, or with film forming protective material dressing to modify the dissolution characteristics of this tablet.Said composition can also be made chewable tablet, for example uses the material of N.F,USP MANNITOL and so in preparation.

In the time need giving chromone compound, can use base-material commonly used with suppository.Theobroma oil is normally used suppository base, can add wax it is modified to improve its melting point a little.Be extensive use of the water miscibility suppository base, especially various molecular weight polyethylene glycol.

The effect that can postpone or prolong described chromone compound by suitable prescription.For example, can prepare the slow molten pill of chromone compound and it is incorporated in tablet or the capsule, or as the slowly-releasing implantable device.This technology also comprises the pill for preparing different dissolution rates and with the mixture filled capsules of this pill.Tablet or capsule can be used undissolved film dressing in the given time.Even parenteral administration also can be made into long-acting form, and method is described chromone compound to be dissolved in or to be suspended in can make it in serum slowly in dispersive oiliness or the emulsification vehicle.

5. embodiment

Following examples are used for illustrative purposes, do not produce any restriction.

5.1 embodiment 1: synthetic 7-hydroxyl-6-methyl-4-(4-(2-tetramethyleneimine-1-base-oxyethyl group)-benzyl)-3-(4-trifluoromethyl)-chromen-2-one

A.1-(2-hydroxyl-4-p-methoxy-phenyl)-2-(4-hydroxy phenyl)-ethyl ketone

(44.69kg is 360mol) with 4-hydroxyphenyl acetic acid (68.5kg, 450mol) suspension in the 144L chlorobenzene with nitrogen purge 3-methoxyphenol.Adding boron trifluoride Anaesthetie Ether compound under 20-25 ℃ (177L, 1440mol).Described suspension is heated to 80 ℃, and stirred 4-5 hour, be cooled to 5-10 ℃ then, and stir and spend the night.

Under nitrogen pressure, filter out sedimentary red/orange solids (unwanted isomer), and filtrate be poured into make its rapid cooling in ice/water.Gained filter cake CH ₂Cl ₂Washing, and slowly add 80%Na ₂CO ₃(aqueous solution) reaches 6-7 up to the pH of the aqueous solution, thereby the boron trifluoride etherate in the solution is cooled off rapidly.Observation gas changes, and required product deposits from solution, forms orange suspension.Subsequent filtration is spent the night in the stirring under 20 ℃ of described orange suspension.Gained filter cake water and methyl tertiary butyl ether (" MTBE ") washing, and dried overnight obtain required product (38kg, 42% productive rate, HPLC purity 95.1% a/a) thus. ¹H NMR(300MHz，DMSO-d ₆)12.30(s，1H)，9.31(s，1H)，7.99(d，1H，J＝9.1Hz)，7.08(d，2H，J＝8.4Hz)，6.70，(d，2H，J＝8.4Hz)，6.53(dd，1H，J＝2.5，9.1Hz)，6.47(d，1H，J＝2.5Hz)，4.18(s，2H)，3.81(s，3H)。MS(ESI)m/z259(M+H) ⁺。

B.4-(4-hydroxybenzyl)-7-methoxyl group-3-(4-trifluoromethyl-phenyl)-chromen-2-one

(13.2g 82mmol) divides 4-trifluoromethylbenzene guanidine-acetic acid (15.2g, 74.45mmol) solution in 120mL DMF under 25 ℃ of several partially disposed with phosphinylidyne diimidazole (" CDI ") in 5 minutes.Described reaction mixture is warming up to 40 ℃, kept 10 minutes, be cooled to room temperature then.Add 1-(2-hydroxyl-4-p-methoxy-phenyl)-2-(4-hydroxy phenyl) ethyl-1-ketone (9.81g, 38mmol), K ₂CO ₃(15.7g, 114mmol) and DMAP (0.93g 7.6mmol), and is warming up to 80 ℃ with described reaction mixture, keeps 2 hours.

Gained suspension is cooled to room temperature, and adds the water of 200mL.Use CH ₂Cl ₂Extract described water layer, and dry described organic layer (MgSO ₄), under vacuum, concentrate then.The gained solid uses purified by flash chromatography (CH ₂Cl ₂: EtOAc), thereby obtain required product (10.2g, 63%).

¹H NMR(300MHz，DMSO-d ₆)9.29(s，1H)，7.79(d，2H，J＝8.7Hz)，7.57(d，2H，J＝8.7Hz)，7.53(d，1H，J＝8.5Hz)，7.04(d，1H，J＝2.3Hz)，6.93(d，2H，J＝8.9Hz)，6.87(dd，1H，J＝8.5，2.3Hz)，6.61(d，2H，J＝8.9Hz)，3.90(s，2H)，3.84(s，3H)。

MS(ESI)m/z 427(M+H) ⁺。

C.4-(7-methoxyl group-2-oxygen-3-(4-trifluoromethyl-phenyl)-2H-chromene-4-base-methyl)-phenyl 2,2-dimethyl-propionic ester

With 4-(4-hydroxybenzyl)-7-methoxyl group-3-(4-trifluoromethyl-phenyl)-chromen-2-one (800mg, 1.88mmol), pivalyl chloride (577 μ L, 4.69mmol), imidazoles (268mg, 3.94mmol) and DMAP (344mg, 2.81mmol) mixture in DMF (10mL) at room temperature stirred 18 hours.Add again part imidazoles (268mg, 3.94mmol), DMAP (344mg, 2.81mmol) and pivalyl chloride (577 μ L 4.69mmol), continue stirring 20 hours again, and total reaction time is 38 hours.Remove described solvent in a vacuum, the gained residue is dissolved among the AcOEt, and use H ₂O washs organic layer.Extract described water layer with AcOEt (2x), and with salt water washing blended organic layer, dry (MgSO ₄) and under vacuum, remove and desolvate.The gained residue use flash chromatography (silica gel, 5: 1 hexanes: AcOEt) purifying, thus make the title compound (820mg, 86%) of white solid. ¹HNMR (300MHz, CDCl ₃) δ 7.64 (d, J=8.0Hz, 2H), 7.43-7.37 (m, 3H), 7.08-7.02 (m, 2H), 7.00-6.93 (m, 2H), 6.88 (d, J=2.5Hz, 1H), 6.77 (dd, J=2.5,8.8Hz, 1H), 4.04 (s, 2H), 3.87 (s, 3H), 1.34 (s, 9H); MS (ESI) m/z 511.0 (M+H) ⁺HPLC t _R7.58 minute.

D.4-(6-bromo-7-methoxyl group-2-oxygen-3-(4-trifluoromethyl-phenyl)-2H-chromene-4-base-methyl)-phenyl 2,2-dimethyl-propionic ester

At room temperature stir 4-(7-methoxyl group-2-oxygen-3-(4-trifluoromethyl-phenyl)-2H-chromene-4-base-methyl)-phenyl 2 from step C, and 2-dimethyl-propionic ester (820mg, 1.61mmol) and Br ₂(83 μ L, 1.61mmol) mixture in AcOH (15mL) is 16 hours.Add the Br of a part in addition ₂(80 μ L 1.55mmol), and continue to stir 6 hours.Described mixture is poured in the water, and used CH ₂Cl ₂(2x) extract water layer.With the described blended organic layer of salt water washing, dry (MgSO ₄) and under vacuum, concentrate.From CH ₂Cl ₂The described residual orange solids of/hexane crystallization, thus make required compound, be white solid (710mg, 75%). ¹H NMR (300MHz, CDCl ₃) δ 7.67 (s, 1H), 7.65 (d, J=8.5Hz, 2H), 7.38 (d, J=8.5Hz, 2H), 7.07-6.94 (m, 4H), 6.89 (s, 1H), 4.01 (s, 2H), 3.98 (s, 3H), 1.35 (s, 9H); MS (ESI) m/z 588.8 (M+H) ⁺HPLCt _R7.78 minute.

E.4-(7-methoxyl group-6-methyl-2-oxygen-3-(4-trifluoromethyl-phenyl)-2H-chromene-4-ylmethyl)-phenyl 2,2-dimethyl-propionic ester

Under reflux temperature, stir 4-(6-bromo-7-methoxyl group-2-oxygen-3-(4-trifluoromethyl-phenyl)-2H-chromene-4-base-methyl)-phenyl 2, and 2-dimethyl-propionic ester (100mg, 0.17mmol), K ₂CO ₃(70mg, 0.51mmol), the trimethylammonium boroxine (24 μ L, 0.17mmol) and Pd (Ph ₃P) ₄(20mg, 0.017mmol) mixture in no Shui diox (2mL) is 2 hours.After being cooled to temperature, gained suspension being filtered on diatomite, and under vacuum condition, remove and desolvate.From CH ₂Cl ₂/ hexane crystalline residue, thus make required compound, be beige solid (72mg, 82%). ¹H NMR (300MHz, CDCl ₃) δ 7.62 (d, J=8.0Hz, 2H), 7.36 (d, J=8.0Hz, 2H), 7.22 (d, J=0.9Hz, 1H), 7.04 (d, J=8.8Hz, 2H), 6.96 (d, J=8.8Hz, 2H), 6.83 (s, 1H), 4.02 (s, 2H), 3.91 (s, 3H), 2.16 (s, 3H), 1.35 (s, 9H); MS (ESI) m/z 524.9 (M+H) ⁺, HPLC tR 7.81 minutes.

F.4-(4-hydroxybenzyl)-7-methoxyl group-6-methyl-3-(4-trifluoromethyl-phenyl)-chromen-2-one

Stir down 4-(7-methoxyl group-6-methyl-2-oxygen-3-(4-trifluoromethyl-phenyl)-2H-chromene-4-base-methyl)-phenyl 2 at 30 ℃, and 2-dimethyl-propionic ester (72mg, 0.14mmol) and LiOH-H ₂(23mg is 0.55mmol) at 8: 1 THF: H for O ₂Mixture in the O mixture (4.5mL) 4 hours.Under vacuum, remove THF, and add AcOEt.The aqueous hydrochloric acid of adding 1N is reduced to 5 with the pH of water layer.Separate each layer, and extract water with AcOEt (2x).With the described blended organic layer of salt water washing, dry (MgSO ₄) and under vacuum, concentrate.Use flash chromatography (99: 1CH ₂Cl ₂: MeOH) the described residue of purifying, thus make required compound, be beige solid (40mg, 67%). ¹H NMR (300MHz, CDCl ₃) δ 7.62 (d, J=8.0Hz, 2H), 7.38 (d, J=8.0Hz, 2H), 7.25 (d, J=1.0Hz, 1H), 6.92-6.86 (m, 2H), 6.82 (s, 1H), 6.75-6.69 (m, 2H), 3.95 (s, 2H), 3.90 (s, 3H), 2.16 (s, 3H); MS (ESI) m/z 440.9 (M+H) ⁺HPLC t _R6.50 minute.

G.7-methoxyl group-6-methyl-4-(4-(2-tetramethyleneimine-1-base-oxyethyl group)-benzyl)-3-(4-trifluoromethyl-phenyl)-chromen-2-one

55 ℃ stir down 4-(4-hydroxybenzyl)-7-methoxyl group-6-methyl-3-(4-trifluoromethyl-phenyl)-chromen-2-ones (50mg, 0.113mmol), K ₂CO ₃(48mg, 0.35mmol) and 1-(2-chloroethyl) tetramethyleneimine hydrochloric acid (24mg, 0.14mmol) mixture in the EtOH (1mL) of moisture (100 μ L) is 5 hours.Described mixture is cooled to room temperature, and is poured into CH ₂Cl ₂-H ₂Among the O.Separate each layer, and use CH ₂Cl ₂(3x) extract water.Dry (MgSO ₄) described blended organic layer, and remove in a vacuum and desolvate.Use flash chromatography (silica gel, 40: 1CH ₂Cl ₂: MeOH) the described residue of purifying, thus make title compound, be beige solid (37mg, 61%).MS (ESI) m/z 537.9 (M+H) ⁺HPLC tR 5.22 minutes.

H.7-hydroxyl-6-methyl-4-(4-(2-tetramethyleneimine-1-base-oxyethyl group)-benzyl)-3-(4-trifluoromethyl-phenyl)-chromen-2-one

130 ℃ down heating 7-methoxyl group-6-methyl-4-(4-(2-tetramethyleneimine-1-base-oxyethyl group)-benzyl)-3-(4-trifluoromethyl-phenyl)-chromen-2-ones (50mg is 0.093mmol) at 1: 1 48%HBr: the solution among the AcOH (3mL) 7 hours.Described mixture is cooled to room temperature, and is poured among the AcOEt/1MNaOH.Separate each layer, and use H ₂O (2x) and salt water washing organic phase, and dry (MgSO ₄).Under vacuum, remove and desolvate.Use the described residue of reversed-phase HPLC purifying, use AcOEt/NaHCO afterwards ₃Extract, thereby make required compound, be yellow solid (25mg, 52%). ¹H NMR (300MHz, CDCl ₃) δ 7.60 (d, J=8.0Hz, 2H), 7.32 (d, J=8.0Hz, 2H), 7.21 (d, J=0.75Hz, 1H), 6.86 (d, J=8.8Hz, 2H), 6.72 (s, 1H), 6.62 (d, J=8.8Hz, 2H), 4.14 (t, J=5.5Hz, 2H), 3.92 (s, 2H), 3.13 (t, J=5.5Hz, 2H), 2.97-2.85 (br s, 4H), 2.13 (s, 3H), 2.00-1.89 (m, 4H); MS (ESI) m/z 523.9 (M) ⁺HPLC t _R5.07 minute.

5.2 embodiment 2: synthetic 3-(2,4 dichloro benzene base)-7-hydroxyl-6-methyl-4-(4-(2-tetramethyleneimine-1-base-oxyethyl group)-benzyl)-chromen-2-one

Except replace 4-(trifluoromethyl) phenylacetic acid among the step B with the 2,4 dichloro benzene guanidine-acetic acid, carry out the synthetic of above-mentioned title compound according to the scheme of embodiment 1.

¹H NMR (300MHz, DMSO) δ 10.8-10-4 (br s, 1H), 7.70 (d, J=2.0Hz, 1H), 7.46 (dd, J=2.0,8.0Hz, 1H), 7.42 (s, 1H), 7.38 (d, J=8.5Hz, 1H), 6.98 (d, J=8.8Hz, 2H), 6.78 (s, 1H), 6.76 (d, J=8.8Hz, 2H), 4.02 (d, J=15.5Hz, 1H), 3.97 (t, J=6.0Hz, 2H), 3.70 (d, J=15.5Hz, 1H), 2.78 (t, J=6.0Hz, 2H), 2.58-2.46 (m, 4H), 2.07 (s, 3H), 1.72-1.61 (m, 4H); MS (ESI) m/z523.8 (M+H) ⁺HPLC t _R5.14 minute.

5.3 embodiment 3: synthetic 3-(2,4 dichloro benzene base)-7-hydroxyl-8-methyl-4-(4-(2-tetramethyleneimine-1-base-oxyethyl group)-benzyl)-chromen-2-one

A.1-(2-hydroxyl-4-p-methoxy-phenyl)-2-(4-hydroxy phenyl)-ethyl ketone

(44.69kg is 360mol) with 4-hydroxyphenyl acetic acid (68.5kg, 450mol) suspension in the 144L chlorobenzene with nitrogen purge 3-methoxyphenol.Adding boron trifluoride Anaesthetie Ether compound under 20-25 ℃ (177L, 1440mol).Gained suspension is heated to 80 ℃, stirred 4-5 hour, be cooled to 5-10 ℃ and stir and to spend the night.

Under nitrogen pressure, filter gained red/orange solids (unwanted isomer) settling, and make its rapid cooling in ice/water by filtrate is poured into.Use CH ₂Cl ₂Washing contains the filter cake of unwanted isomer, and slowly adds 80%Na ₂CO ₃(aqueous solution) reaches 6-7 up to the pH of the aqueous solution, thereby the boron trifluoride etherate in the solution is cooled off rapidly.Observation gas changes, and required product is come out from solution deposition, forms orange suspension.The stirring under 20 ℃ of described orange suspension is spent the night, filter then.Gained filter cake water and MTBE washing, and dried overnight, thus required product (38kg, 42% productive rate, HPLC purity 95.1% a/a) obtained. ¹H NMR(300MHz，DMSO-d ₆)12.30(s，1H)，9.31(s，1H)，7.99(d，1H，J＝9.1Hz)，7.08(d，2H，J＝8.4Hz)，6.70，(d，2H，J＝8.4Hz)，6.53(dd，1H，J＝2.5，9.1Hz)，6.47(d，1H，J＝2.5Hz)，4.18(s，2H)，3.81(s，3H)。MS(ESI)m/z 259(M+H) ⁺。

B.3-(2,4 dichloro benzene base)-4-(4-hydroxybenzyl)-7-methoxyl group-chromen-2-one

Except using 2,4 dichloro benzene guanidine-acetic acid replacement 4-trifluoromethylbenzene guanidine-acetic acid, prepare above-mentioned title compound as the described method of embodiment 1B.20g ketone (77.5mmol) and 31.6g acid (155mmol) produce 27.52g product (83%). ¹H NMR(300MHz，DMSO-d ₆)9.26(s，1H)，7.58(d，1H，J＝8.8Hz)，7.50(dd，1H，J＝1.9，8.2Hz)，7.45(d，1H，J＝8.2Hz)，7.06(d，1H，J＝2.2Hz)，6.90(d，3H，J＝8.2Hz)，6.59(d，2H，J＝8.2Hz)，3.98(d，1H，J＝15.4Hz)，3.85(s，3H)，3.69(d，1H，J＝15.4Hz)。MS(ESI)m/z428(M+H) ⁺。

C.4-(4-(2-bromine oxethyl)-benzyl)-3-(2,4 dichloro benzene base)-7-methoxyl group-chromen-2-one

Under reflux temperature, stir 3-(2,4 dichloro benzene base)-4-(4-hydroxybenzyl)-7-methoxyl group-chromen-2-one (6.4g, 15.0mmol), glycol dibromide (12.9mL, 149.8mmol) and K ₂CO ₃(4.14g, 30mmol) mixture in acetone (65mL) is 20.5 hours.The K that in this time, adds another part ₂CO ₃(241.6mg, 1.75mmol) and glycol dibromide (3.2mL, 37.5mmol), restir is 7.5 hours afterwards.Continue heating 20 hours again, total reaction time is 48 hours.The gained mixture is cooled to room temperature, and is poured into CH ₂Cl ₂/ H ₂Among the O.Separate each layer, and use CH ₂Cl ₂(2x) extract water.With salt solution (2x) washing blended organic phase, and dry (MgSO ₄), and under vacuum, remove and desolvate.The gained residue uses flash chromatography (300g silica gel, 4.5: 1: 1 → 4: 1: 1 → 3: 1: 1 hexane: CH ₂Cl ₂: AcOEt) carry out purifying, thereby make title compound, be white solid (4.851g, 61%), and the raw material (2.054g, 32%) that reclaims. ¹H NMR (300MHz, CDCl ₃) δ 7.50 (d, J=2.0Hz, 1H), 7.44 (d, J=9.0Hz, 1H), 7.26 (dd, J=2.0,8.0Hz, 1H), 7.12 (d, J=8.0Hz, 1H), 6.98-6.91 (m, 2H), 6.88 (d, J=2.0Hz, 1H), 6.79-6.74 (m, 3H), 4.23 (t, J=6.5Hz, 2H), 4.04 (d, J=15.5Hz, 1H), 3.87 (s, 3H), 3.78 (d, J=15.5Hz, 1H), 3.61 (t, J=6.5Hz, 2H); MS (ESI) m/z 532.7 (M+H) ⁺HPLCt _R8.38 minute.

D.4-(4-(2-bromine oxethyl)-benzyl)-3-(2,4 dichloro benzene base)-7-hydroxyl-chromen-2-one

At room temperature stir 4-(4-(2-bromine oxethyl)-benzyl)-3-(2,4 dichloro benzene base)-7-methoxyl group-chromen-2-one (4.0g, 7.49mmol), AlCl ₃(5.89g, 44.2mmol) and EtSH (2.77mL is 37.4mmol) at CH ₂Cl ₂Mixture (200mL) 2.5 hours.Slowly pour described mixture into CH ₂Cl ₂/ saturated NaHCO ₃In the aqueous solution.Separate each layer, use CH again ₂Cl ₂(3x) extract water.Make the pH of water layer rise to 10 with the 2MNaOH aqueous solution, and extract with AcOEt (3x).With each organic layer of salt water washing, and dry (MgSO ₄), under vacuum condition, concentrate described blended organic phase.The gained residue uses flash chromatography to carry out purifying (250g silica gel, 95: 5 → 9: 1CH ₂Cl ₂: AcOEt), thereby make title compound, separate obtaining Off-white solid (3.454g, 89%). ¹H NMR (300MHz, CDCl ₃) δ 7.50 (d, J=2.0Hz, 1H), 7.42 (d, J=8.8Hz, 1H), 7.25 (dd, J=2.0,8.0Hz, 1H), 7.12 (d, J=8.0Hz, 1H), 6.99 (d, J=2.0Hz, 1H), 6.94 (d, J=8.5Hz, 2H), 6.78 (d, J=8.5Hz, 2H), 6.73 (dd, J=2.0,8.8Hz, 1H), 6.61-6.49 (the bandwidth signals peak, 1H), 4.23 (t, J=6.0Hz, 2H), 4.04 (d, J=15.5Hz, 1H), 3.78 (d, J=15.5Hz, 1H), 3.61 (t, J=6.0Hz, 2H); MS (ESI) m/z 516.9 (M-M) ^-HPLCt _R7.41 minute.

E.4-(4-(2-bromine oxethyl)-benzyl)-3-(2,4 dichloro benzene base)-7-hydroxyl-8-iodo-chromen-2-one

(4-(2-bromine oxethyl)-benzyl)-(2.734g is 5.26mmol) at NH for 3-(2,4 dichloro benzene base)-7-hydroxyl-chromen-2-one toward 4- ₄OH (22mL), MeOH (55mL) and CH ₂Cl ₂Add I in the solution (33mL) ₂(1.334g 5.26mmol), and at room temperature stirred the mixture obtain 30 minutes.Add the 4MHCl aqueous solution then, the pH of water layer is reduced to 5.Separate each layer, and extract water with AcOEt (3x).With salt solution (2x) washing blended organic layer, dry (MgSO ₄) and concentrate in a vacuum.Gained beige solid (3.25g, 96%) is directly used in next step, need not to be further purified. ¹H NMR (300MHz, DMSO) δ 11.43 (s, 1H), 7.74 (d, J=2.0Hz, 1H), and 7.51-7.42 (m, 3H), 7.02 (d, J=8.5Hz, 2H), 6.82 (d, J=8.8Hz, 1H), 6.78 (d, J=8.5Hz, 2H), 4.21 (t, J=5.0Hz, 2H), 4.04 (d, J=15.5Hz, 1H), 3.73 (t, J=5.0Hz, 2H), 3.70 (d, J=15.5Hz, 1H); MS (ESI) m/z 642.7 (M-H) ^-HPLC t _R7.27 minute.

F.3-(2,4 dichloro benzene base)-7-hydroxyl-8-iodo-4-(4-(2-tetramethyleneimine-1-base-oxyethyl group)-benzyl)-chromen-2-one

Toward 4-(4-(2-bromine oxethyl)-benzyl)-3-(2, the 4-dichlorophenyl)-7-hydroxyl-8-iodo-chromen-2-one (1.90g, 2.94mmol) add tetramethyleneimine (1.46mL in the suspension of stirring in anhydrous THF (20mL), 17.6mmol), the gained yellow solution is heated to 85 ℃, kept 1.5 hours.The gained mixture is cooled to room temperature, and under vacuum condition, removes and desolvate.The gained residue is dissolved in CH ₂Cl ₂In, and use H ₂O and salt water washing, dry (MgSO ₄).Concentrate described organic phase in a vacuum.The gained residue is used flash chromatography (200g silica gel, 12: 1 → 9: 1 → 85: 15CH ₂Cl ₂: MeOH) purifying, thus make title compound, be yellow solid (1.87g, 100%). ¹H NMR (300MHz, CDCl ₃) δ 7.50 (d, J=2.0Hz, 1H), 7.32 (d, J=8.8Hz, 1H), 7.26 (dd, J=2.0,8.0Hz, 1H), 7.12 (d, J=8.0Hz, 1H), and 6.95-6.86 (m, 3H), 6.77-6.70 (m, 2H), 4.30 (t, J=5.0Hz, 2H), 4.00 (d, J=15.5Hz, 1H), 3.76 (d, J=15.5Hz, 1H), 3.25 (t, J=5.0Hz, 2H), and 3.19-3.04 (m, 4H), 2.04-1.91 (m, 4H); MS (ESI) m/z 635.6 (M) ⁺HPLC t _R5.85 minute.

G.3-(2,4 dichloro benzene base)-7-hydroxyl-8-methyl-4-(4-(2-tetramethyleneimine-1-base-oxyethyl group)-benzyl)-chromen-2-one

Under reflux temperature, stir 3-(2,4 dichloro benzene base)-7-hydroxyl-8-iodo-4-(4-(2-tetramethyleneimine-1-base-oxyethyl group)-benzyl)-chromen-2-one (1.85g, 2.91mmol), K ₂CO ₃(1.21g, 8.75mmol), the trimethylammonium boroxine (427 μ L, 3.05mmol) and Pd (Ph ₃P) ₄(336mg, 0.29mmol) mixture in no Shui diox (22mL) is 2.5 hours.After the gained green suspension is cooled to room temperature, under vacuum condition, removes and desolvate.The gained residue is dissolved in CH ₂Cl ₂In, and water and salt water washing organic layer, dry (MgSO ₄), and concentrate in a vacuum.The gained residue uses flash chromatography (125g silica gel, 8: 0.5: 0.5 → 7: 0.5: 0.5CH ₂Cl ₂: MeOH: AcOEt → 9: 1CH ₂Cl ₂: MeOH) purifying, thus yellow solid obtained, and described solid (is used CH afterwards with reversed-phase HPLC ₂Cl ₂/ NaHCO ₃Extract) be further purified.Separate obtaining title compound, be yellow solid (220.4mg, 14%). ¹HNMR (300MHz, CDCl ₃) δ 7.48 (d, J=2.0Hz, 1H), 7.24 (dd, J=2.0,8.0Hz, 1H), 7.18 (d, J=8.8Hz, 1H), 7.12 (d, J=8.0Hz, 1H), 6.90 (d, J=8.8Hz, 2H), 6.84 (d, J=8.8Hz, 1H), 6.72 (d, J=8.8Hz, 2H), 4.23 (t, J=5.0Hz, 2H), 4.00 (d, J=15.5Hz, 1H), 3.74 (d, J=15.5Hz, 1H), 3.24 (t, J=5.0Hz, 2H), 3.15-3.04 (m, 4H), 2.20 (s, 3H), 2.05-1.93 (m, 4H); MS (ESI) m/z 523.8 (M) ⁺HPLC t _R4.74 minute.

5.4 embodiment 4: synthetic 8-ethanoyl-3-(2,4 dichloro benzene base)-7-hydroxyl-4-(4-(2-tetramethyleneimine-1-base-oxyethyl group)-benzyl)-chromen-2-one

Embodiment 3 is described as mentioned carries out steps A-F, and step G replaces with following step H.

H.

Under reflux temperature, stir 3-(2, the 4-dichlorophenyl)-7-hydroxyl-8-iodo-4-(4-(2-tetramethyleneimine-1-base-oxyethyl group)-benzyl)-chromen-2-one (187mg, 0.294mmol), tributyl (1-vinyl ethyl ether base) tin (119 μ L, 0.353mmol) and Pd (Ph ₃P) ₄(34mg, 0.0294mmol) mixture in no Shui diox (6mL) is 2 hours.After being cooled to room temperature, adding the 1M HCl aqueous solution, and resulting mixture was stirred 20 minutes.Add CH ₂Cl ₂, and separate each layer.Use CH again ₂Cl ₂(2x) extract water layer.Dry (MgSO ₄) blended organic layer and concentrated in a vacuum.The gained residue uses flash chromatography (silica gel, 19: 1 → 9: 1CH ₂Cl ₂: MeOH) carry out purifying, thereby obtain filbert solid (48mg, HPLC purity 84%), this solid is carried out recrystallization and makes title compound, be yellow solid (22mg, 14%) with the AcOEt/ hexane.

I.

The product of step H is dissolved in the mixture of 2-propyl alcohol/concentrated HCI.After at room temperature stirring 20 minutes, evaporating solvent.With the gained solid at Et ₂Smash to pieces among the O, and filter and make the 12mg title compound, be the form of its hydrochloride. ¹H NMR (300MHz, DMSO) δ 11.57 (s, 1H), 10.05-9.91 (bandwidth signals peak, 1H), 7.72 (d, J=2.0Hz, 1H), 7.56 (d, J=8.8Hz, 1H), 7.48 (dd, J=2.0,8.0Hz, 1H), 7.44 (d, J=8.0Hz, 1H), 7.04 (d, J=8.5Hz, 2H), and 6.86-6.78 (m, 3H), 4.20 (t, J=4.5Hz, 2H), 4.02 (d, J=15.5Hz, 1H), 3.73 (d, J=15.5Hz, 1H), 3.60-3.45 (m, 4H), 3.25-2.97 (m, 2H), 2.58 (s, 3H), 2.05-1.75 (m, 4H); MS (ESI) m/z 552.0 (M+H) ⁺HPLC t _R5.33 minute.

5.5 embodiment 5: synthetic 3-(4-chloro-phenyl-)-7-hydroxyl-8-iodo-4-{ (4-(2-piperidyl oxyethyl group) phenyl) methyl } the 2H-chromen-2-one

Toward 3-(4-chloro-phenyl-)-7-hydroxyl-4-{ (4-(2-piperidyl oxyethyl group) phenyl) methyl }-add ammonium hydroxide (3mL) and unit price iodine (0.259g) in methyl alcohol (10mL) solution of 2H-chromen-2-one (0.500g).This reaction mixture was at room temperature stirred 10 minutes.Under reduced pressure, remove and desolvate.With methylene dichloride (3X) and the resulting solid of water extraction.Organic layer is mixed, and use dried over mgso.Filter out sal epsom, and under reduced pressure, remove and desolvate, make yellow solid.By preparation HPLC (20-50% acetonitrile/water, 20mL/ minute) the described solid of purifying, thereby obtain title compound (0.290g, 46% productive rate). ¹H-NMR(CDCl ₃)δ7.34(dd，3H)，7.20(d，2H)，6.92(d，2H)，6.84(d，1H)，6.75(d，2H)，4.13(t，2H)，3.97(s，2H)，2.84(t，2H)，2.61(bs，3H)，1.643(m，4H)，1.46(m，2H)，1.24(m，1H)。MS(m/z)616(M+1) ⁺。

5.6 embodiment 6: synthetic 3-(4-chloro-phenyl-)-7-hydroxyl-2-oxygen-4-{ (4-(2-piperidyl oxyethyl group) phenyl)-2H-chromene-8-nitrile-2-ketone

Toward 3-(4-chloro-phenyl-)-7-hydroxyl-8-iodo-4-{ (4-(2-piperidyl oxyethyl group) phenyl) methyl }-2H-chromen-2-one (embodiment 5 described making as mentioned) (1.50g) adds cupric cyanide (1.05g) in the solution of dimethyl formamide (30mL).Described solution was heating 18 hours in the screw thread mouth flask under 120 ℃.After being cooled to room temperature, described solvent is removed under reduced pressure.Resulting oil methylene dichloride and water extraction.Mix organic layer and use dried over mgso.Filter out this sal epsom, and under reduced pressure, remove and desolvate.Described thick solid passes through preparation HPLC (30-65% acetonitrile/water, 20mL/ minute, 30 minutes) purifying, thereby obtains title compound (0.125g, 10% productive rate).MS(m/z)515(M+1) ⁺。

5.7 embodiment 7: synthetic 7-hydroxyl-8-third-2-thiazolinyl-4-{ (4-(2-pyrrolidyl oxyethyl group) phenyl) methyl }-3-(4-(trifluoromethyl) phenyl)-2H-chromen-2-one

A.7-third-2-alkene oxygen base-4-{ (4-(2-pyrrolidyl oxyethyl group) phenyl) methyl }-3-(4-(trifluoromethyl)-2H-chromen-2-one

Toward 7-hydroxyl-4-{ (4-(2-pyrrolidyl oxyethyl group) phenyl) methyl }-add allyl alcohol (0.226mL) and triphenylphosphine (0.641g) in 3-(4-(trifluoromethyl) the phenyl)-solution of 2H-chromen-2-one (0.550g) in tetrahydrofuran (THF) (6mL) and methylene dichloride (6mL).Then, di-isopropyl azodicarboxy hydrochlorate (0.437mL) is added drop-wise in this solution.Gained mixture stir about 1 hour (or intact) up to raw materials consumption.Under reduced pressure, concentrate described solution, and with methylene dichloride (3X) and water extraction.Described organic layer carries out drying with sal epsom, filters out this sal epsom, and concentrates described solvent under reduced pressure, thereby obtains orange oil.Described oil carries out purifying by silica gel chromatography (6-10% ethanol/methylene), thereby make 7-third-2-alkene oxygen base-4-{ (4-(2-pyrrolidyl oxyethyl group) phenyl) methyl }-3-(4-(trifluoromethyl) phenyl)-2H-chromen-2-one (0.245g, 40% productive rate).MS(m/z)550(M+1) ⁺。

B.7-hydroxyl-8-third-2-thiazolinyl-4-{ (4-(2-pyrrolidyl oxyethyl group) phenyl) methyl }-3-(4-(trifluoromethyl) phenyl-2H-chromen-2-one

With 7-third-2-alkene oxygen base-4-{ (4-(2-pyrrolidyl oxyethyl group) phenyl) methyl }-3-(4-(trifluoromethyl) phenyl)-2H-chromen-2-one (0.150g) is dissolved in the Diethyl Aniline (4mL), and stirred 2 hours down at 190 ℃.Gained solution methylene dichloride (3X) and 1M hydrochloric acid extraction.Gained organic layer dried over sodium sulfate, and filter, described solvent is removed under reduced pressure, thereby makes rough brown solid.Described solid obtains title compound (0.027g, 18% productive rate) with preparation HPLC (40-100% acetonitrile/water, 20mL/ minute) purifying. ¹H-NMR(CDCl ₃)δ7.63(d，2H)，7.38(d，2H)，7.25(d，1H)，6.92(d，2H)，6.75(d，2H)，6.70(d，1H)，6.0(m，1H)，5.13(dd，2H)，4.12(t，2H)，3.94(s，2H)，3.66(d，2H)，3.05(bs，2H)，2.8(bs，4H)，1.25(s，2H)。MS(m/z)550(M+1) ⁺。

5.8 embodiment 8: synthetic 8-((dimethylamino) methyl)-3-(4-chloro-phenyl-) 7-hydroxyl-4-{ ((2-piperidyl oxyethyl group) phenyl) methyl }-the 2H-chromen-2-one

In water (0.414mL) solution of 37% formaldehyde, add 2.0M xylidine (2.5mL).At room temperature stirred this solution 15 minutes, and it joined 7-hydroxyl-4-{ (4-(the 2-pyrrolidyl oxyethyl group) phenyl) methyl that is dissolved in methyl alcohol (5mL) and the methylene dichloride (2mL) }-3-(4-(trifluoromethyl) phenyl)-2H-chromen-2-one (0.250g) in.With resulting mixture heating up to 70 ℃, keep about 3 hours (but or till can not detecting the raw material of detection limit) by tlc.Described solvent is removed under reduced pressure, and gained oil extracts with methylene dichloride (3X) and sodium hydrogen carbonate solution.Described organic layer is mixed and use dried over mgso, filter out this sal epsom, and under reduced pressure except that desolvating.Gained oil passes through preparation HPLC (20-50% acetonitrile/water, 20mL/ minute, 30 minutes) purifying, thereby makes title compound (0.130g., 46% productive rate). ¹H-NMR(CDCl ₃)δ7.33(dd，3H)，7.18(d，2H)，6.93(d，2H)，6.78(d，2H)，6.75(d，1H)，4.05(t，3H)，3.94(s，2H)，2.74(t，2H)，2.48(bs，3H)，2.42(s，6H)，1.59(m，4H)，1.44(m，2H)，1.25(m，1H)。MS(m/z)547(M+1) ⁺。

5.9 embodiment 9:6-bromo-7-hydroxyl-4-(4-(2-tetramethyleneimine-1-base-oxyethyl group)-benzyl)-3-(4-trifluoromethyl-phenyl)-chromen-2-one

A.1-(5-bromo-2-hydroxyl-4-methoxyl group-phenyl)-2-(4-hydroxyl-phenyl)-ethyl ketone

(3.6mL, acetate 75.6mmol) (50mL) solution stirring is added drop-wise to 1-(2-hydroxyl-4-methoxyl group-phenyl)-2-(4-hydroxyl-phenyl)-ethyl ketone, and (20g in acetate 77.4mmol) (500mL) solution, stirs this reaction mixture 8 hours with bromine.Then, under reduced pressure, remove acetate, with resulting solid and from methanol recrystallization, and make the 8.9g crude product.Use anti-phase preparation HPLC (the 30-100% acetonitrile+0.1% TFA aqueous solution+0.1% TFA carried out 18 minutes, and 100% acetonitrile carried out 15 minutes then) to be further purified, make level part, this grade part is neutralized with sodium bicarbonate, and use ethyl acetate extraction.The organic grade of part that obtains is mixed, and, remove under the reduced pressure and desolvate, thereby make title compound (4.8g, 18%) by dried over mgso and filtration.MS(ESI)m/z337.3(M+1) ⁺，339.3(M+1+2) ⁺。

B.6-bromo-4-(4-hydroxyl-benzyl)-7-methoxyl group-3-(4-trifluoromethyl-phenyl)-chromen-2-one

In 80 ℃ oil bath, heat 1-(5-bromo-2-hydroxyl-4-methoxyl group-phenyl)-2-(4-hydroxyl-phenyl)-ethyl ketone (0.64g, 1.9mmol), 4-trifluoromethylbenzene guanidine-acetic acid (0.78g, 3.8mmol), phosphinylidyne diimidazole (0.62g, 3.8mmol), N, N-dimethyl aminopyridine (0.12g, 1.0mmol) and salt of wormwood (0.52g, 3.8mmol) mixture overnight in DMF (15mL).Cooled this reaction mixture is poured in the water, and extracts with EtOAc.Organic extraction is mixed, and water and the continuous mutually washing of saturated sodium-chloride, use dried over mgso, filter and under reduced pressure, concentrate.Resulting material is with silica gel flash column chromatography (1: 2EtOAc: hexane) carry out purifying, thereby make title compound (0.82g, 85%).MS(ESI)m/z 505.0(M+1) ⁺，507.0(M+1+2) ⁺。

C.6-bromo-7-methoxyl group-4-(4-(2-tetramethyleneimine-1-base-oxyethyl group)-benzyl)-3-(4-trifluoromethyl-phenyl)-chromen-2-one

To 6-bromo-4-(4-hydroxyl-benzyl)-7-methoxyl group-3-(4-trifluoromethyl-phenyl)-chromen-2-one (0.70g, 1.4mmol) and triphenylphosphine (0.73g, 2.8mmol) add 1-(2-hydroxyethyl) tetramethyleneimine (0.65mL successively in the solution in THF (40mL), 5.6mmol) and the di-isopropyl azo-2-carboxylic acid (0.55mL, 2.8mmol).At room temperature stirring this reaction mixture spends the night.This reaction mixture is poured in the water, and extracted with EtOAc.Organic extraction is mixed and water and the continuous mutually washing of saturated sodium-chloride, use dried over mgso, filter and under reduced pressure, concentrate.(10% methyl alcohol is at CH to use the silica gel flash column chromatography ₂Cl ₂In) purifying, make solid title compound (0.44g, 53%).MS(ESI)m/z602.3(M+1) ⁺，604.3(M+1+2) ⁺。

D.6-bromo-7-hydroxyl-4-(4-(2-tetramethyleneimine-1-base-oxyethyl group)-benzyl)-3-(4-trifluoromethyl-phenyl)-chromen-2-one

(0.20g 0.3mmol) is heated to backflow with the solution of hydrogen bromide in acetate (10mL) and spends the night with 6-bromo-7-methoxyl group-4-(4-(2-tetramethyleneimine-1-base-oxyethyl group)-benzyl)-3-(4-trifluoromethyl-phenyl)-chromen-2-one.After cooling, excessive acetate is removed in decompression down.The residue of gained is dissolved in ethyl acetate, and washs with saturated sodium bicarbonate, water and saturated sodium-chloride are continuous mutually; Use dried over mgso, filter and under reduced pressure, concentrate, thereby make crude product.Use anti-phase preparation HPLC (the 20-80% acetonitrile+0.1% TFA aqueous solution+0.1% TFA carried out 30 minutes) to carry out purifying, make level part, this grade part is neutralized with sodium bicarbonate subsequently, and use ethyl acetate extraction.Organic moiety is mixed and use dried over mgso, filter and under reduced pressure concentrated, thereby make title compound (＞98% purity, 45.0mg, 23%). ¹H NMR(300MHz，d ₆-DMSO)δ7.78(m，2H)，7.69(s，1H)，7.55(m，2H)，7.08(m，2H)，6.88(s，1H)，6.83(m，2H)，4.04(t，2H)，3.94(s，2H)，2.92(t，2H)，2.67(bm，4H)，1.72(bm，4H)。MS(ESI)m/z 588.3(M+1) ⁺，590.3(M+1+2) ⁺。

Embodiment 10

Other representational chromone compound

Following table 1 discloses representational chromone compound.These chromone compounds can use method as herein described to make.

Table 1

Representational chromone compound

And pharmacy acceptable salt, in the formula:

No.	R ₁	R ₂	X	Y	n
No.	R ₁	R ₂	X	Y	n	1	H	C(＝O)CH ₃	Cl	Cl	1
2	CH ₃	H	Cl	Cl	1	1	H	C(＝O)CH ₃	Cl	Cl	1
2	CH ₃	H	Cl	Cl	1	3	H	CH ₃	Cl	Cl	1
4	H	CH ₃	H	CF ₃	1	3	H	CH ₃	Cl	Cl	1
4	H	CH ₃	H	CF ₃	1	5	CH ₃	H	H	CF ₃	1
6	C(＝O)CH ₃	H	H	CF ₃	1	5	CH ₃	H	H	CF ₃	1
6	C(＝O)CH ₃	H	H	CF ₃	1	7	H	C(＝O)CH ₃	H	CF ₃	1
8	H	CN	H	CF ₃	1	7	H	C(＝O)CH ₃	H	CF ₃	1
8	H	CN	H	CF ₃	1	9	Br	H	H	CF ₃	1
10	H	I	H	Cl	2	9	Br	H	H	CF ₃	1
10	H	I	H	Cl	2	11	-CH(OH)CH ₃	H	CF ₃	H	1

Embodiment 11

Suppress the release of IL-6

Tested exemplary chromone compound and suppressed the ability that people U-2OS osteosarcoma cell discharges IL-6, this osteosarcoma cell personnel selection ER-α or ER-β stable transfection (Stein, B.; Yang, M.X.Mol.Cell.Biol 15:4971-4979,1995; Poli, V. etc., EMBO J.13:1189-1196,1994).But the IL-6 that measures the parental generation untransfected U-2OS clone of the ER-α do not express detection level or ER-β discharges in contrast.IC ₅₀The chromone compound of＜100nM is a bone resorption inhibitor in the useful especially body.Therefore, this chromone compound especially can be used for treating osteoporosis, Paget's disease and metastatic bone cancer.These chromone compounds also can be used as anticancer preparation, are some cancers because the IL-6 level raises, as multiple myeloma, prostate cancer, ovarian cancer, the rectum cancer and cervical cancer pathogenic because of.

Adopt standard molecular biological technique, the expression vector stable transfection people U-2OS osteosarcoma cell (ATCC) of personnel selection total length ER-α or ER-β.Obtain expressing the stable subclone of high-level ER-α or ER-β mRNA.Expression with RNA enzyme protection analysis verification ER-α or ER-β.But parental generation U-2OS cell is not expressed the ER-α or the ER-β of any detection limit.

Cell is joined in the no phenol red medium in 96 orifice plates with the density of 80,000 cells in every hole, and this cultivation contains the foetal calf serum through activated carbon adsorption.Handle cell with vehicle (0.2% DMSO) or chromone compound (0.01-1000nm is with 0.2% DMSO preparation) after 24 hours.Use 2.5ng/ml TNF α and 1ng/ml IL-1 β irritation cell after 30 minutes.Analyze the output of the cytokine (IL-6) in the substratum supernatant after 24 hours according to the explanation of manufacturers with commercially available ELISA test kit.Cytokine production when having vehicle (0.2% DMSO) is made as 100%.

Result's (table 2) uses IC ₅₀(nM) value representation, the amount of the IL-6 that produces when this value representation exists with respect to vehicle are enough to suppress the concentration of the chromone compound that 50% IL-6 produces.The result shows that exemplary chromone compound has suppressed the release of IL-6, therefore can be used for treatment or prevention bone-resorbing disease.

Embodiment 12

Suppress MCF-7 breast cancer cell propagation

This embodiment has shown the ability of exemplary chromone compound in vitro inhibition MCF-7 breast cancer cell 17 beta estradiol dependencys growths, and has compared them and with reference to the activity of SERM.The MCF-7 cell is the fabulous vitro system of research compound to the effect of estrogen-dependent growth of breast cancers.(May，F.E.B.；Westley，B.R.J.Biol.Chem.262：15894-15899，1987)。IC ₅₀The chromone compound of＜100nM is especially effectively to resist-the mammary cancer agent in the body.

With the MCF-7 breast cancer cell with every hole 5 * 10 ³The density of individual cell joins in 24 orifice plates in no phenol red DMEM: F-12 (1: the 1) substratum, and this substratum contains 1% microbiotic, 0.05% mercaptoethanol, 0.01% thanomin, 0.42ng/mL Sodium Selenite and 5% FCS through activated carbon adsorption.

In the MCF-7 breast cancer cell of cultivating, add exemplary chromone compound (0.1-1000nM is with 0.2% DMSO preparation) and 0.1nM 17 beta estradiols, kept 72 hours.Add then ³The thymus pyrimidine of H-mark is also measured its incorporation in cell at incubation after 4 hours.Result's (table 2) uses IC ₅₀(nM) value representation, this value representation are enough to suppress the concentration of the chromone compound of 50% MCF-7 breast cancer cell growth with respect to contrast.The result shows that exemplary chromone compound has suppressed MCF-7 breast cancer cell propagation, the therefore mammary cancer that can be used for treating or preventing the patient.

Table 2

Vitro data

Therefore, as above shown in the table 2, embodiment 11 and 12 in vitro results have shown that described chromone compound can be used for treatment or prevention bone-resorbing disease and cancer.

Embodiment 13

Suppress BG-1 ovarian cancer cell propagation

This mensuration is used for confirming the ability of exemplary chromone compound in the growth of vitro inhibition BG-1 ovarian cancer cell 17 beta estradiol dependencys, and has compared them and with reference to the activity of SERM.The BG-1 cell is to estimate the effective external model (Greenberger, L.M. etc., Clin.Cancer Res.7:3166-3177,2001) of estrogen antagonist compound to the effect of ovarian tumor growth.

With the BG-1 ovarian cancer cell with every hole 5 * 10 ³The density of individual cell joins in the no phenol red DMEM in 24 orifice plates: F-12 (1: the 1) substratum, and this substratum contains 1% microbiotic, 0.05% mercaptoethanol, 0.01% thanomin, 0.42ng/mL Sodium Selenite and 5% FCS through activated carbon adsorption.In the BG-1 ovarian cancer cell of cultivating, add exemplary chromone compound (0.1-1000nM prepares with 0.2%DMSO) and 0.1nM 17 beta estradiols and cultivated 72 hours.Add afterwards ³The thymus pyrimidine of H-mark is also measured its incorporation in cell at incubation after 4 hours.Result IC ₅₀(nM) value representation, this value representation are enough to suppress the concentration of the chromone compound of 50% BG-1 ovarian cancer cell growth with respect to contrast.Exemplary chromone compound has suppressed the growth of BG-1 ovarian cancer cell, therefore the ovarian cancer that can be used for treating or preventing the patient.

Embodiment 14

Rat pharmacokinetics (PK) is analyzed

The standard test of P of Rats K boxlike

Gavage by per os gives exemplary chromone compound and the internal standard substance raloxifene that the rat dosage level is 5 mg/kg body weight.Blood sampling in 15 minutes to 24 hours the time durations after administration.Blood sample is passed through acetonitrile precipitation, centrifugal, and in vacuum centrifuge, evaporate supernatant.The exsiccant resistates is dissolved in the methanol (60: 40v/v) and that contains 1% formic acid at UPTISPHERETM C18 reversed-phase HPLC post (particle diameter: 3 μ m; The post specification: 2 * 50mm) upward analyze by HPLC.Elutriant A is 10% acetonitrile solution (pH 2.1) that contains 0.1% formic acid, and elutriant B is 90% acetonitrile (pH 2.1) that contains 10% water and 0.1% formic acid.With 5-100%B linear elution 7 minutes, kept 3 minutes at 100%B then, column temperature is 50 ℃ of constant temperature.Constant flow rate is 0.4mL/min.The application of sample volume is 10 μ L.The effluent liquid of HPLC system is directly imported in the ion source of Agilent 1100 serial MS detectors (single quadrupole mass spectrometer) and and carry out normal atmosphere electrospray ionization (holotype) it.All compounds are as protonated quasi-molecular ion (M+H) ⁺Detect.Raloxifene is with the internal standard substance that performs an analysis.Based on the blood levels that the 7-horizontal alignment curve (in triplicate) that adopts blank rat blood sample comes the quantization ratio compound, added external standard and interior mark storage liquid in this blank rat blood sample.

The checking of P of Rats K boxlike

Raloxifene (3 mg/kg) is distinguished per os (p.o.) separately give 4 female rats.By collection blood sample mentioned above and analyzing.The data of this checking research pharmacokinetic data that produces and the raloxifene that obtains in the boxlike dosetest are compared to detect possible pharmacokinetics reaction.The deviation (be about each parameter peaked ± 50%) that exceeds the canonical biometric disparity range is thought to take place between the compound in the box indication of pharmacokinetics reaction strongly, and each this data are abandoned.

Embodiment 15

The pharmacokinetics feature of exemplary chromone compound in stump-tailed macaque after intravenously and the oral administration

Dosage intravenously with 0.5 mg/kg gives to give chromone compound with the oral dose of 1.0 mg/kg.For intravenous administration, the 10mg compound is dissolved in the 0.4mL 1-Methyl-2-Pyrrolidone and transfers to 2mL with PEG 200, gained concentration is 5mg/mL.

With chromone compound as the suspension orally give.This suspension makes by the 20mg compound is dissolved in about 18mL solution, and this solution is to be dissolved in 1.0% hydroxypropylcellulose (HPC, 2.5% (w/v) oxysuccinic acid w/v).Afterwards, with 1N NaOH pH is transferred to 4.5 and volume transferred to 20mL.The ultimate density that obtains test compounds is 1.0mg/mL.

With the chromone compound intravenous injection to saphena (volume injected is 0.1mL/kg, is about 30 seconds inject time).Oral administration carries out (the administration volume is 1mL/kg, with the flushing of 10ml water) by the gavage of per os.Be given 2 animals (to intravenously and oral, n=2) for every kind of mode of administration chromone compound.

Pierce through from the beginning vein or saphena is collected blood sample (the about 0.5ml of each time point) by vein, place the test tube that is coated with EDTA, test tube is placed on ice immediately and keep in cold storage at-20 ℃.

Test according to following time scheme:

(a) the be untreated blood sample (time 0, baseline) of animal;

(b) give test compounds; With

(c) the 5th, 15,30,60,120,180,240,420 minute, 24 and 48 hours blood sample (intravenously and oral administration) after the administration.

Table 3

Zoologize and pharmacological agent

Animal	Treatment	Dosage	Approach
Animal	Treatment	Dosage	Approach	1 2 3 4	Chromone compound chromone compound chromone compound chromone compound	0.5 mg/kg 0.5 mg/kg 1.0 mg/kg 1.0 mg/kg	The intravenously intravenously is oral

The scope of the specific embodiments that the invention is not restricted among the embodiment to be disclosed, these embodiments be just in order to illustrate aspects more of the present invention, any in embodiment identical on the function all within the scope of the invention.In fact, except above show and describe those, various modifications of the present invention are conspicuous and within the scope of the appended claims for being proficient in those skilled in the art.

Quoted many bibliographys, its complete content is included in this paper as a reference in full.

Claims

1. compound or its pharmacy acceptable salt with following structure:

In the formula:

X and Y are hydrogen, halogen or (halo) (C independently _1-6Alkyl);

N is 1,2 or 3; With

One of following situation:

(c) R ₁And R ₂Be-CH ₃

2. compound as claimed in claim 1 or its pharmacy acceptable salt, wherein this compound has following structure:

3. one kind prepares the compound with following structure or the method for its pharmacy acceptable salt:

This method comprises makes the compound with following structure or the step of its pharmacy acceptable salt demethylation:

In the formula:

X and Y are hydrogen, halogen or (halo) (C independently _1-6Alkyl);

N is 1,2 or 3; With

One of following situation:

(c) R ₁And R ₂Be-CH ₃

4. the method for treatment or prevention bone-resorbing disease, this method comprises the compound as claimed in claim 1 that needs the patient of this treatment or prevention significant quantity or the pharmacy acceptable salt of this compound.

5. method as claimed in claim 4, wherein said bone-resorbing disease are that osteolytic lesion, Paget's disease or the bone relevant with hyperparathyroidism that osteoporosis, metastatic bone cancer, hypercalcemia, orthopaedic implants cause runs off.

6. the treatment or the method for prophylaxis of tumours disease, this method comprise the compound as claimed in claim 1 that needs the patient of this treatment or prevention significant quantity or the pharmacy acceptable salt of this compound.

7. method as claimed in claim 6, wherein said neoplastic disease is a cancer.

8. method as claimed in claim 7, wherein said cancer are head and neck, eye, skin, mouth, larynx, esophagus, chest, bone, blood, lung, colon, sigmoid colon, rectum, stomach, prostate gland, mammary gland, ovary, uterus, kidney, liver, pancreas, brain, intestines, heart or adrenal cancer.

9. method as claimed in claim 7, wherein said cancer are the cancers of uterus, ovary or mammary gland.

10. method that activates the function of ER in the osteocyte, this method comprise the compound as claimed in claim 1 that needs the patient of this treatment or prevention significant quantity or the pharmacy acceptable salt of this compound.

11. method as claimed in claim 10, wherein said cell is an osteosarcoma cell.

12. a method that suppresses the function of ER in breast cancer cell, ovarian cancer cell, endometrial carcinoma cell, uterus carcinoma cell, prostate cancer cell or the hypothalamus cancer cells, this method comprise the compound as claimed in claim 1 that makes described cells contacting significant quantity or the pharmacy acceptable salt of this compound.

13. one kind is suppressed the method that IL-6 expresses, this method comprises the compound as claimed in claim 1 of the cells contacting significant quantity that allows to express ER and IL-6 or the pharmacy acceptable salt of this compound.

14. method as claimed in claim 13, wherein said cell is an osteocyte.

15. a method that suppresses to express the growth of tumour cell of ER, this method comprise the compound as claimed in claim 1 of the tumour cell contact significant quantity that allows to express ER or the pharmacy acceptable salt of this compound.

16. method as claimed in claim 15, wherein said cell is a cancer cells.

17. the method for the disease that treatment or prevention worsen when oestrogenic hormon exists, this method comprise the compound as claimed in claim 1 that needs the patient of this treatment or prevention significant quantity or the pharmacy acceptable salt of this compound.

18. method as claimed in claim 17, wherein said disease is a cancer.

19. the method for the disease that treatment or prevention improve when oestrogenic hormon exists, this method comprise the compound as claimed in claim 1 that needs the patient of this treatment or prevention significant quantity or the pharmacy acceptable salt of this compound.

20. method as claimed in claim 19, wherein said disease is a bone-resorbing disease.

21. vascular protection, CNS effect, acne or cataractous method after a treatment or prevention endometriosis, hypercholesterolemia, prostatomegaly, prostate cancer, obesity, flush, skin effect, anxious state of mind, memory loss, menopausal syndrome, alopecia (baldness), type ii diabetes, degenerative brain disorder, the urinary incontinence, gastrointestinal tract disease, the damage, this method comprise the compound as claimed in claim 1 that needs the patient of this treatment or prevention significant quantity or the pharmacy acceptable salt of this compound.

22. a treatment or preventing cardiovascular disease, hirsutism, multiple myeloma or lymphadenomatous method, this method comprise the compound as claimed in claim 1 that needs the patient of this treatment or prevention significant quantity or the pharmacy acceptable salt of this compound.

23. the spermatogenetic method of prevention, this method comprises the compound as claimed in claim 1 that needs the patient of this prevention significant quantity or the pharmacy acceptable salt of this compound.

24. the method for the bad reproduction influence that a prevention is relevant with being exposed to the imbalance of environmental chemistry or natural hormone, this method comprise the compound as claimed in claim 1 that needs the patient of this prevention significant quantity or the pharmacy acceptable salt of this compound.

25. compound as claimed in claim 1 or the pharmacy acceptable salt of this compound and composition of pharmaceutically acceptable carrier or vehicle that contains significant quantity.